TW202411244A - GLP-1/GIP dual agonist, and preparation method therefor and use thereof - Google Patents

Abstract

A long-acting GLP-1/GIP dual agonist compound, wherein the compound has dual agonistic effects on a glucagon-like peptide-1 (GLP-1) receptor and a human glucose-dependent insulinotropic polypeptide (GIP) receptor. The present invention also relates to the use of the compound in the preparation of a drug for treating diabetes.

Description

GLP-1/GIP dual agonist and its preparation method and use

The present invention belongs to the field of medicine, and specifically relates to a GLP-1/GIP dual agonist and its use in preparing a drug for treating diabetes.

Type II diabetes is a chronic metabolic disease caused by insulin resistance and characterized by high blood sugar. Elevated blood sugar can increase the incidence and mortality of various other diseases. Common treatments such as increased exercise, controlled diet, oral hypoglycemic drugs, and injected therapeutic drugs can only temporarily maintain blood sugar balance, but cannot fundamentally treat diabetes and the many complex complications caused by high blood sugar. Therefore, it is of great clinical significance to actively explore and develop diabetes treatment drugs with new hypoglycemic mechanisms and high safety.

Glucagon-like peptide-1 (GLP-1) is a hormone mainly produced by intestinal L cells and belongs to the incretin group. GLP-1 is an important intestinal insulinotropic hormone secreted by intestinal L cells. Its main physiological effects in the body include stimulating the secretion and release of insulin, inhibiting the secretion of glucagon, promoting the proliferation of pancreatic β cells and inhibiting their apoptosis, inhibiting gastric emptying, and promoting the production of satiety. Glucose-dependent insulinotropic polypeptide (GIP), which regulates blood sugar through insulin and glucagon, has also been widely confirmed and applied.

Compounds with GLP-1/GIP dual agonist activity have been described in WO2013164483A1, WO2014192284A1, and WO2016111971A1.

Fatty acid modification of peptides can, for example, extend the half-life through the physical binding motif of their fatty acid chains to the albumin subdomain, thereby improving the pharmacokinetics of peptides. Although the use of fatty acids can improve the half-life of peptides, the specific effect of the extension and some side effects are still technical issues that need to be solved.

A range of side effects associated with GLP-1 include: For example, since animal studies and drug database analyses have shown that GLP-1 receptor agonists are associated with pancreatitis, pancreatic cancer, and thyroid cancer, there are concerns that these drugs will cause effects on pancreatic and thyroid tissue. Other case reports have linked the use of GLP-1 (mainly exenatide) to the occurrence of acute kidney injury, mainly due to hemodynamic disturbances caused by nausea, vomiting, and diarrhea. The most common symptoms associated with the use of GLP-1 receptor agonists are gastrointestinal symptoms, mainly nausea. Other common adverse reactions include injection site reactions, headache, and nasopharyngitis. (Filippatos TD, Panagiotopoulou TV, Elisaf MS. Adverse Effects of GLP-1 Receptor Agonists. Rev Diabet Stud. 2014 Fall-Winter;11(3-4):202-30. doi: 10.1900/RDS.2014.11.202. Epub 2015 Feb 10. PMID: 26177483; PMCID: PMC5397288.)

Regarding the side effects of tirzepatide (Mathiesen DS, Bagger JI, Bergmann NC, Lund A, Christensen MB, Vilsbøll T, Knop FK. The Effects of Dual GLP-1/GIP Receptor Agonism on Glucagon Secretion-A Review. Int J Mol Sci. 2019 Aug 22;20(17):4092. doi: 10.3390/ijms20174092. PMID: 31443356; PMCID: PMC6747202.): At the highest dose, the number of adverse events with tirzepatide exceeded that with dulaglutide, especially gastrointestinal adverse events (66.0% for patients taking 15 mg tirzepatide and 2.3% for patients taking 1.5 mg The researchers found that LY3298176 had a higher incidence of hypoglycemia in patients taking tirzepatide (42.6%) and a higher incidence of hypoglycemia in patients taking dulaglutide (7.5% of participants taking 15 mg tirzepatide and 3.7% of participants taking dulaglutide) (Frias J.P., Nauck M.A., Van J., Kutner M.E., Cui X., Benson C., Urva S., Gimeno R.E., Milicevic Z., Robins D., et al. Efficacy and safety of LY3298176, a novel dual GIP and GLP-1 receptor agonist, in patients with type 2 diabetes: a randomised, placebo-controlled and active comparator-controlled phase 2 trial. Lancet. 2018;392:2180–2193. doi: 10.1016/S0140-6736(18)32260-8. ). To circumvent this problem, a titration strategy to reduce adverse events of tirzepatide has been reported (Frias J.P., Nauck M.A., Van J., Benson C., Bray R., Milicevic Z., Haupt A., Robins D.A. 993-P: A 12-week, randomized, placebo-controlled study assessing the efficacy and safety of three dose-escalation algorithms of tirzepatide, a novel dual GIP and GLP-1 receptor agonist, in patients with type 2 diabetes. Diabetes. 2019;68:993. doi: 10.2337/db19-993-P. ), and a three-step dose-escalation regimen of 15 mg tirzepatide did not appear to be effective in reducing the overall incidence of gastrointestinal side effects compared with previously reported studies (Frias J.P., Nauck M.A., Van J., Kutner M.E., Cui X., Benson C., Urva S., Gimeno R.E., Milicevic Z., Robins D., et al. Efficacy and safety of LY3298176, a novel dual GIP and GLP-1 receptor agonist, in patients with type 2 diabetes: a randomised, placebo-controlled and active comparator-controlled phase 2 trial. Lancet. 2018;392:2180–2193. doi: 10.1016/S0140-6736(18)32260-8. ). The incidence of hypoglycemic episodes also increased to ~16% in participants taking 15 mg of tilbotide. Although telbociclib treatment is still associated with promising weight loss (about 5.6 kg after 12 weeks), the high incidence of gastrointestinal side effects does bring some uncertainty to the clinical potential of telbociclib. In addition, the titration strategy in the telbociclib marketing instructions is time-consuming, with an initial dose of 2.5 mg, which is increased to 5 mg after 4 weeks. If an increase in dose is needed to control blood sugar, the current dose must be maintained for 4 weeks and then increased by 2.5 mg, which is very inconvenient.

There is still a medical need to provide new compounds that are dual agonists of GIP and GLP-1 receptors and support the possibility of once-weekly dosing in humans, providing options with better safety, better efficacy, and more convenient medication.

One object of the present invention is to provide some long-acting GLP-1/GIP dual agonist compounds. Compared with the GIP/GLP-1 dual agonist compounds in the field, the compounds of the present invention have a longer half-life and/or a more sustained duration of action. The compounds of the present invention have a stronger hypoglycemic effect. The compounds of the present invention have lower side effects, including but not limited to gastrointestinal reactions and cardiac safety risks. The compounds of the present invention can adopt a shorter titration strategy. The compounds of the present invention have a larger safety window. In addition, the compounds of the present invention have a protective effect on the liver.

As part of the present invention, the applicant discovered that the selection of the length, composition and attachment position of the fatty acid chain, as well as the linker between the peptide and the fatty acid chain, has an unexpected effect on the extension of the half-life of the polypeptide.

One embodiment of the present invention is to provide a compound of formula (AI) and a pharmaceutically acceptable salt thereof: YX ₁ -EGTX ₂ -TSDY-A11-A12-A13-LDK-A17-AQ-A20-EFVKWLLK-A29-GPSSGAPPPSK Formula (AI); wherein X ₁ is Aib; X ₂ is αMePhe; A11 is Aib or Ala; A12 is Ala, Ile, Lys, Phe or Pya(4); A13 is Aib, Cha, Leu, αMePhe or αMeTyr; A17 is Gln or Ile; A20 is Ala or Ser; A29 is Gln or Gly; 1 K, 2 K, 3 K or 4 K positions are selected from among the 16th, 24th, 28th and 40th K positions, and are chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -Z to the ε-amine group of the K side chain, wherein each a is independently an integer from 0 to 5, each b is independently an integer from 0 to 5, each c is independently an integer from 10 to 24, and Z is independently selected from -CH ₃ , carboxylic acid or carboxylic acid bioisostere, phosphate/ester or sulfonate/ester.

In another embodiment, the present invention provides a compound of formula (AI) or a pharmaceutically acceptable salt thereof, wherein, _X1 is Aib; _X2 is αMePhe; A11 is Aib or Ala; A12 is Ala, Ile, Lys, Phe or Pya(4); A13 is Aib, Cha, Leu, αMePhe or αMeTyr; A17 is Gln or Ile; A20 is Ala or Ser; A29 is Gln or Gly, and 1, 2, 3 or 4 K positions are selected from among positions 16, 24, 28 and 40, by using ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -Z is chemically modified by binding to the ε-amine group of the K side chain, wherein a is independently selected from an integer of 1, 2, 3, 4 or 5, b is independently selected from an integer of 1, 2, 3, 4 or 5, and c is independently selected from an integer of 12 to 22; wherein Z is independently selected from -CH ₃ , a carboxylic acid or a carboxylic acid bioisostere, a phosphate/ester or a sulfonate/ester; and further provided is a compound wherein c is independently selected from 14, 16, 18 or 20.

The peptide of the above formula (AI) or a pharmaceutically acceptable salt thereof, wherein A11 is Aib; or/and The peptide of the above formula (AI) or a pharmaceutically acceptable salt thereof, wherein A12 is Ile; or/and The peptide of the above formula (AI) or a pharmaceutically acceptable salt thereof, wherein A13 is Aib; or/and The peptide of the above formula (AI) or a pharmaceutically acceptable salt thereof, wherein A17 is Gln; or/and The peptide of the above formula (AI) or a pharmaceutically acceptable salt thereof, wherein A20 is Ala; or/and The peptide of the above formula (AI) or a pharmaceutically acceptable salt thereof, wherein A29 is Gly.

One embodiment of the present invention is to provide a compound of formula (I) and a pharmaceutically acceptable salt thereof: _YX1 - _EGTX2 - _TSDYX3 - _IX4 -LDKQAQAEFVKWLLKGGPSSG-APPPSK Formula (I); wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; 1, 2, 3 or 4 K positions are selected from positions 16, 24, 28 and 40, by using ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a- (γ-Glu) _b -CO-( _CH2 ) _c -Z is chemically modified by binding to the ε-amine group of the K side chain, wherein each a is independently an integer from 0 to 5, each b is independently an integer from 0 to 5, each c is independently an integer from 10 to 24, wherein Z is independently selected from -CH ₃ , carboxylic acid or carboxylic acid bioisostere, phosphate/ester or sulfonate/ester; and the C-terminal amino acid is amidated to form a C-terminal primary amide.

In some embodiments, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein one K position is selected from among positions 16, 24, 28, and 40, and is chemically modified by binding ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -Z to the ε-amine group of the K side chain. For example, at the 16-position K position, the ε-amine group of the K side chain is chemically modified by binding with ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -Z; for example, at the 24-position K position, the ε-amine group of the K side chain is chemically modified by binding with ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -Z; for example, at the 28-position K position, the ε-amine group of the K side chain is chemically modified by binding with ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -Z is chemically modified by binding to the ε-amine group of the K side chain; for example, at the 40-position K, it is chemically modified by binding to the ε-amine group of the K side chain with ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -Z.

In some embodiments, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein two K positions are selected from positions 16, 24, 28, and 40, and two modified chains are chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -Z to the ε-amine group of the K side chain. For example, at the positions of K at the 16th position and K at the 24th position, two modification chains are chemically modified by binding to the ε-amine group of the K side chain with ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -Z; for example, at the positions of K at the 16th position and K at the 28th position, two modification chains are chemically modified by binding to the ε-amine group of the K side chain with ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -Z; for example, at the positions of K at the 16th position and K at the 40th position, two modification chains are chemically modified by binding to the ε-amine group of the K side chain with ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -Z is bound to the ε-amine group of the K side chain to chemically modify two modification chains; for example, at the 24-position K and 28-position K positions, ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -Z is bound to the ε-amine group of the K side chain to chemically modify two modification chains; for example, at the 24-position K and 40-position K positions, ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -Z is attached to the ε-amine group of the K side chain to carry out chemical modification of two modification chains; for example, at the positions of K 28 and K 40, two modification chains are chemically modified by attaching ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -Z to the ε-amine group of the K side chain.

In some embodiments, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein three K positions are selected from positions 16, 24, 28, and 40, and three modification chains are chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -Z to the ε-amine group of the K side chain. For example, at the positions of K at the 16th position, K at the 24th position, and K at the 28th position, three modification chains are chemically modified by binding to the ε-amine group of the K side chain with ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -Z; for example, at the positions of K at the 16th position, K at the 24th position, and K at the 40th position, three modification chains are chemically modified by binding to the ε-amine group of the K side chain with ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -Z; for example, at the positions of K at the 16th position, K at the 28th position, and K at the 40th position, three modification chains are chemically modified by binding to the ε-amine group of the K side chain with ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -Z is linked to the ε-amine group of the K side chain to carry out chemical modification of three modification chains; for example, at the positions of K 24, K 28 and K 40, three modification chains are chemically modified by linking ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -Z to the ε-amine group of the K side chain.

In some embodiments, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein four modification chains are chemically modified by binding ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -Z to the ε-amine group of the K side chain at four positions selected from among positions 16, 24, 28, and 40. For example, four modification chains are chemically modified by binding ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -Z to the ε-amine group of the K side chain at positions 16, 24, 28, and 40.

In some embodiments, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein Z is independently selected from -CH ₃ , carboxylic acid or carboxylic acid bioisostere, phosphate/ester or sulfonate/ester, for example, but not limited to, the Z may include carboxylic acid (-CO ₂ H) or carboxylic acid bioisostere (e.g. ), phosphoric acid (-P(O)(OH) ₂ ) or sulfonic acid (-SO ₂ OH) group, preferably -CO ₂ H.

Carboxylic acid bioisosteres, suitable carboxylic acid bioisosteres are known in the art. Preferably, the bioisostere has a proton with a _pKa similar to that of the corresponding carboxylic acid. Examples of suitable bioisosteres may include, but are not limited to, tetrazoles, acylsulfonamides, acylhydroxylamides, and squaric acid derivatives, as shown below: , , , , R is Me or CF ₃ .

In some embodiments, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein a is independently selected from an integer of 1, 2, 3, 4 or 5, preferably 1, 2 or 3, b is independently selected from an integer of 1, 2, 3, 4 or 5, preferably 1, 2 or 3, and c is independently selected from an integer of 12 to 22; in addition, the present invention provides a compound wherein c is 14, 16, 18 or 20.

One embodiment of the present invention is to provide a compound of formula (I) and a pharmaceutically acceptable salt thereof: _YX1 - _EGTX2 - _TSDYX3 - _IX4 -LDKQAQAEFVKWLLKGGPSSG-APPPSK Formula (I); wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; one or two K positions selected from positions 16, 24, 28, and 40 are chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a- (γ-Glu) _b -CO-( _CH2 ) _c -Z to the ε-amine group of the K side chain, wherein Z is independently selected from _-CH3 , carboxylic acid or carboxylic acid bioisostere, phosphate/ester or sulfonate/ester, preferably _-CO2 H, each a is independently an integer from 0 to 5, each b is independently an integer from 0 to 5, and each c is independently an integer from 10 to 24; and the C-terminal amino acid is amidated to form a C-terminal primary amide.

In another embodiment, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; one or two K positions selected from positions 16, 24, 28, and 40 are chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a- (γ-Glu) _b -CO-( _CH2 ) _c -Z to the ε-amine group of the K side chain, wherein Z is independently selected from _-CH3 , carboxylic acid or carboxylic acid bioisostere, phosphate/ester or sulfonate/ester, preferably _-CO2 H, a is independently selected from an integer of 1, 2, 3, 4 or 5, b is independently selected from an integer of 1, 2, 3, 4 or 5, and c is independently selected from an integer of 12 to 22; in addition, the present invention provides a compound wherein c is 14, 16, 18 or 20.

One embodiment of the present invention is to provide a compound of formula (I) and a pharmaceutically acceptable salt thereof: _YX1 - _EGTX2 - _TSDYX3 - _IX4 -LDKQAQAEFVKWLLKGGPSSG-APPPSK Formula (I); wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at position 24 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a- (γ-Glu) _b -CO-( _CH2 ) _c - _CO2H to the ε-amine group of the K side chain, wherein a is independently selected from an integer from 0 to 5, b is independently selected from an integer from 0 to 5, and c is independently selected from an integer from 10 to 24; and the C-terminal amino acid is amidated to form a C-terminal primary amide.

In another embodiment, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at position 24 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a- (γ-Glu) _b -CO-( _CH2 ) _c - _CO2H to the ε-amine group of the K side chain, wherein a is independently selected from 1, 2, 3, 4 or 5 integers, b is independently selected from 1, 2, 3, 4 or 5 integers, and c is independently selected from integers from 12 to 22; the present invention also provides compounds wherein c is 14, 16, 18 or 20.

Another embodiment of the present invention is to provide a compound of formula (I) and a pharmaceutically acceptable salt thereof: _YX1 - _EGTX2 - _TSDYX3 - _IX4 -LDKQAQAEFVKWLLKGGPSSG-APPPSK Formula (I); wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at position 28 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a- (γ-Glu) _b -CO-( _CH2 ) _c -CO2H to the ε-amine group of the K side chain, wherein a is independently selected from an integer from 0 to 5, b is independently selected from an integer from 0 to 5, and c is independently selected from an integer from 10 to 24; and the C _- terminal amino acid is amidated to form a C-terminal primary amide.

In another embodiment, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at position 28 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a- (γ-Glu) _b -CO-( _CH2 ) _c - _CO2H to the ε-amine group of the K side chain, wherein a is independently selected from 1, 2, 3, 4 or 5 integers, b is independently selected from 1, 2, 3, 4 or 5 integers, and c is independently selected from integers from 12 to 22; the present invention also provides a compound wherein c is 14, 16, 18 or 20.

Another embodiment of the present invention is to provide a compound of formula (I) and a pharmaceutically acceptable salt thereof: _YX1 - _EGTX2 - _TSDYX3 - _IX4 -LDKQAQAEFVKWLLKGGPSSG-APPPSK Formula (I); wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at position 16 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a- (γ-Glu) _b -CO-( _CH2 ) _c - _CO2H to the ε-amine group of the K side chain, wherein a is independently selected from an integer from 0 to 5, b is independently selected from an integer from 0 to 5, and c is independently selected from an integer from 10 to 24; and the C-terminal amino acid is amidated to form a C-terminal primary amide.

In another embodiment, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at position 16 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a- (γ-Glu) _b -CO-( _CH2 ) _c - _CO2H to the ε-amine group of the K side chain, wherein a is independently selected from 1, 2, 3, 4 or 5 integers, b is independently selected from 1, 2, 3, 4 or 5 integers, and c is independently selected from integers from 12 to 22; the present invention also provides compounds wherein c is 14, 16, 18 or 20.

Another embodiment of the present invention is to provide a compound of formula (I) and a pharmaceutically acceptable salt thereof: _YX1 - _EGTX2 - _TSDYX3 - _IX4 -LDKQAQAEFVKWLLKGGPSSG-APPPSK Formula (I); wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at position 40 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a- (γ-Glu) _b -CO-( _CH2 ) _c - _CO2H to the ε-amine group of the K side chain, wherein a is independently selected from an integer from 0 to 5, b is independently selected from an integer from 0 to 5, and c is independently selected from an integer from 10 to 24; and the C-terminal amino acid is amidated to form a C-terminal primary amide.

In another embodiment, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at position 40 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a- (γ-Glu) _b -CO-( _CH2 ) _c - _CO2H to the ε-amine group of the K side chain, wherein a is independently selected from 1, 2, 3, 4 or 5 integers, b is independently selected from 1, 2, 3, 4 or 5 integers, and c is independently selected from integers from 12 to 22; the present invention also provides compounds wherein c is 14, 16, 18 or 20.

Another embodiment of the present invention is to provide a compound of formula (I) and a pharmaceutically acceptable salt thereof: _YX1 - _EGTX2 - _TSDYX3 - _IX4 -LDKQAQAEFVKWLLKGGPSSG-APPPSK Formula (I); wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at positions 24 and 28 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a- (γ-Glu) _b -CO-( _CH2 ) _c - _CO2 H is linked to the ε-amine group of the K side chain to chemically modify two modification chains, wherein each a is independently an integer from 0 to 5, each b is independently an integer from 0 to 5, and each c is independently an integer from 10 to 22; and the C-terminal amino acid is amidated to form a C-terminal primary amide.

In another embodiment, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at positions 24 and 28 is chemically modified with two modification chains by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a- (γ-Glu) _b -CO-( _CH2 ) _c - _CO2H to the ε-amine group of the K side chain, wherein a is independently selected from 1, 2, 3, 4 or 5 integers, b is independently selected from 1, 2, 3, 4 or 5 integers, and c is independently selected from integers from 12 to 22; the present invention also provides compounds wherein c is 14, 16, 18 or 20.

Another embodiment of the present invention is to provide a compound of formula (I) and a pharmaceutically acceptable salt thereof: _YX1 - _EGTX2 - _TSDYX3 - _IX4 -LDKQAQAEFVKWLLKGGPSSG-APPPSK Formula (I); wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at positions 16 and 24 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a- (γ-Glu) _b -CO-( _CH2 ) _c - _CO2 H is linked to the ε-amine group of the K side chain to chemically modify two modification chains, wherein each a is independently an integer from 0 to 5, each b is independently an integer from 0 to 5, and each c is independently an integer from 10 to 22; and the C-terminal amino acid is amidated to form a C-terminal primary amide.

In another embodiment, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at positions 16 and 24 is chemically modified with two modification chains by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a- (γ-Glu) _b -CO-( _CH2 ) _c - _CO2H to the ε-amine group of the K side chain, wherein a is independently selected from 1, 2, 3, 4 or 5 integers, b is independently selected from 1, 2, 3, 4 or 5 integers, and c is independently selected from integers from 12 to 22; the present invention also provides compounds wherein c is 14, 16, 18 or 20.

Another embodiment of the present invention is to provide a compound of formula (I) and a pharmaceutically acceptable salt thereof: _YX1 - _EGTX2 - _TSDYX3 - _IX4 -LDKQAQAEFVKWLLKGGPSSG-APPPSK Formula (I); wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at positions 16 and 28 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a- (γ-Glu) _b -CO-( _CH2 ) _c - _CO2 H is linked to the ε-amine group of the K side chain to chemically modify two modification chains, wherein each a is independently an integer from 0 to 5, each b is independently an integer from 0 to 5, and each c is independently an integer from 10 to 22; and the C-terminal amino acid is amidated to form a C-terminal primary amide.

In another embodiment, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at positions 16 and 28 is chemically modified with two modification chains by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a- (γ-Glu) _b -CO-( _CH2 ) _c - _CO2H to the ε-amine group of the K side chain, wherein a is independently selected from 1, 2, 3, 4 or 5 integers, b is independently selected from 1, 2, 3, 4 or 5 integers, and c is independently selected from integers from 12 to 22; the present invention also provides compounds wherein c is 14, 16, 18 or 20.

Another embodiment of the present invention is to provide a compound of formula (I) and a pharmaceutically acceptable salt thereof: _YX1 - _EGTX2 - _TSDYX3 - _IX4 -LDKQAQAEFVKWLLKGGPSSG-APPPSK Formula (I); wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at positions 16 and 40 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a- (γ-Glu) _b -CO-( _CH2 ) _c - _CO2 H is linked to the ε-amine group of the K side chain to chemically modify two modification chains, wherein each a is independently an integer from 0 to 5, each b is independently an integer from 0 to 5, and each c is independently an integer from 10 to 22; and the C-terminal amino acid is amidated to form a C-terminal primary amide.

In another embodiment, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at positions 16 and 40 is chemically modified with two modification chains by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a- (γ-Glu) _b -CO-( _CH2 ) _c - _CO2H to the ε-amine group of the K side chain, wherein a is independently selected from 1, 2, 3, 4 or 5 integers, b is independently selected from 1, 2, 3, 4 or 5 integers, and c is independently selected from integers from 12 to 22; the present invention also provides compounds wherein c is 14, 16, 18 or 20.

Another embodiment of the present invention is to provide a compound of formula (I) and a pharmaceutically acceptable salt thereof: _YX1 - _EGTX2 - _TSDYX3 - _IX4 -LDKQAQAEFVKWLLKGGPSSG-APPPSK Formula (I); wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at positions 24 and 40 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a- (γ-Glu) _b -CO-( _CH2 ) _c - _CO2 H is linked to the ε-amine group of the K side chain to chemically modify two modification chains, wherein each a is independently an integer from 0 to 5, each b is independently an integer from 0 to 5, and each c is independently an integer from 10 to 22; and the C-terminal amino acid is amidated to form a C-terminal primary amide.

In another embodiment, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at positions 24 and 40 is chemically modified with two modification chains by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a- (γ-Glu) _b -CO-( _CH2 ) _c - _CO2H to the ε-amine group of the K side chain, wherein a is independently selected from 1, 2, 3, 4 or 5 integers, b is independently selected from 1, 2, 3, 4 or 5 integers, and c is independently selected from integers from 12 to 22; the present invention also provides compounds wherein c is 14, 16, 18 or 20.

Another embodiment of the present invention is to provide a compound of formula (I) and a pharmaceutically acceptable salt thereof: _YX1 - _EGTX2 - _TSDYX3 - _IX4 -LDKQAQAEFVKWLLKGGPSSG-APPPSK Formula (I); wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at positions 28 and 40 is replaced by ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a- (γ-Glu) _b -CO-( _CH2 ) _c - _CO2 H is linked to the ε-amine group of the K side chain to chemically modify two modification chains, wherein each a is independently an integer from 0 to 5, each b is independently an integer from 0 to 5, and each c is independently an integer from 10 to 22; and the C-terminal amino acid is amidated to form a C-terminal primary amide.

In another embodiment, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at positions 28 and 40 is chemically modified with two modification chains by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a- (γ-Glu) _b -CO-( _CH2 ) _c - _CO2H to the ε-amine group of the K side chain, wherein a is independently selected from 1, 2, 3, 4 or 5 integers, b is independently selected from 1, 2, 3, 4 or 5 integers, and c is independently selected from integers from 12 to 22; the present invention also provides compounds wherein c is 14, 16, 18 or 20.

In all the above embodiments, each a is independently an integer from 1 to 3, each b is independently an integer from 1 to 3, and each c is independently an integer from 12 to 22; in addition, the present invention provides compounds wherein c is 14, 16, 18 or 20. In all the above embodiments, it is preferred that a modification chain is performed on K at position 24 or 28.

In one embodiment, the present invention provides a compound of the following formula or a pharmaceutically acceptable salt thereof; _{YX1- EGTX2} - _TSDYX3 - _IX4- LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at position 24 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _2- (γ-Glu) ₁ -CO-( _CH2 ) ₁₆ - _CO2H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO:7).

In one embodiment, the present invention provides a compound of the following formula or a pharmaceutically acceptable salt thereof; _{YX1- EGTX2} - _TSDYX3 - _IX4- LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at position 24 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _2- (γ-Glu) ₁ -CO-( _CH2 ) ₁₈ - _CO2H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 13).

In one embodiment, the present invention provides a compound of the following formula or a pharmaceutically acceptable salt thereof; _{YX1- EGTX2} - _TSDYX3 - _IX4- LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at position 24 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _2- (γ-Glu) ₁ -CO-( _CH2 ) ₂₀ - _CO2H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 14).

In one embodiment, the present invention provides a compound of the following formula or a pharmaceutically acceptable salt thereof; _{YX1- EGTX2} - _TSDYX3 - _IX4- LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at position 24 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _2- (γ-Glu) ₃ -CO-( _CH2 ) ₁₆ - _CO2H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 15).

In one embodiment, the present invention provides a compound of the following formula or a pharmaceutically acceptable salt thereof; _{YX1- EGTX2} - _TSDYX3 - _IX4- LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at position 24 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _3- (γ-Glu) ₁ -CO-( _CH2 ) ₂₀ - _CO2H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 16).

In one embodiment, the present invention provides a compound of the following formula or a pharmaceutically acceptable salt thereof; _{YX1- EGTX2} - _TSDYX3 - _IX4- LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at position 28 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _2- (γ-Glu) ₁ -CO-( _CH2 ) ₁₆ - _CO2H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO:8).

In one embodiment, the present invention provides a compound of the following formula or a pharmaceutically acceptable salt thereof; _{YX1- EGTX2} - _TSDYX3 - _IX4- LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at position 28 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _2- (γ-Glu) ₁ -CO-( _CH2 ) ₂₀ - _CO2H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 17).

In one embodiment, the present invention provides a compound of the following formula or a pharmaceutically acceptable salt thereof; _{YX1- EGTX2} - _TSDYX3 - _IX4- LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at position 28 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _2- (γ-Glu) ₃ -CO-( _CH2 ) ₁₆ - _CO2H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 18).

In one embodiment, the present invention provides a compound of the following formula or a pharmaceutically acceptable salt thereof; _{YX1- EGTX2} - _TSDYX3 - _IX4- LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at position 28 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _3- (γ-Glu) ₁ -CO-( _CH2 ) ₁₆ - _CO2H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 19).

In one embodiment, the present invention provides a compound of the following formula or a pharmaceutically acceptable salt thereof; _{YX1- EGTX2} - _TSDYX3 - _IX4- LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at position 28 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _3- (γ-Glu) ₁ -CO-( _CH2 ) ₂₀ - _CO2H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO:20).

The present invention also provides a method for increasing the duration of action of a GLP-1/GIP dual agonist in a patient, characterized in that the acylated GLP-1/GIP dual agonist provided by the present invention as described above is used.

In one embodiment, the present invention provides a composition comprising a compound of the present invention and a pharmaceutically acceptable carrier, diluent or excipient.

One embodiment provides the use of a compound according to any one of the above embodiments in the preparation of a medicament for treating or preventing hyperglycemia, impaired glucose tolerance, type I diabetes, type II diabetes, obesity, hypertension, syndrome X, dyslipidemia, cognitive impairment, atherosclerosis, myocardial infarction, coronary heart disease and other cardiovascular diseases, stroke, inflammatory bowel syndrome, dyspepsia and gastric ulcer.

One embodiment provides the use of a compound according to any one of the above embodiments for the preparation of a medicament for delaying or preventing disease progression in type II diabetes.

One embodiment provides the use of a compound according to any one of the above embodiments for the preparation of a medicament for reducing food intake, reducing β-cell apoptosis, increasing β-cell function and β-cell mass and/or restoring β-cell glucose sensitivity.

In one embodiment, the present invention provides a method for treating type II diabetes, comprising administering an effective amount of a compound of the present invention to a patient in need thereof.

In one embodiment, the present invention provides a method for improving glycemic control in a patient with type II diabetes, comprising administering to a patient in need thereof an effective amount of a compound of the present invention as an adjunct to diet and exercise.

In one embodiment, the present invention provides a method for long-term weight management in a patient with an initial body mass index ≥ 27 and type II diabetes, comprising administering to the patient in need thereof an effective amount of a compound of the present invention as an adjunct to a hypocaloric diet and increased physical activity.

In one embodiment, the present invention provides a method for treating metabolic syndrome, comprising administering an effective amount of a compound of the present invention to a patient in need thereof. In another embodiment, the present invention provides a method for treating dyslipidemia, obesity and/or hepatic steatosis associated with insulin resistance and diabetes, comprising administering an effective amount of a compound of the present invention to a patient in need thereof. In addition, the present invention provides a method for treating weakness or increasing bone strength, comprising administering an effective amount of a compound of the present invention to a patient in need thereof.

In one embodiment, the present invention provides compounds of the present invention as an adjunct to diet and exercise for glycemic control in patients with type II diabetes. In one embodiment, the present invention provides compounds of the present invention as an adjunct to a hypocaloric diet and increased physical activity for long-term weight management in patients with an initial body mass index ≥ 27 and type II diabetes.

The compounds of the present invention can be used as drugs for treating or preventing various diseases, including obesity, due to the above-mentioned activation of GLP-1 receptors and GIP receptors. The compounds of the present invention can be used as drugs for preventing or treating the following diseases: for example, symptomatic obesity, obesity based on simple obesity, disease states or diseases related to obesity, eating disorders, diabetes (for example, type I diabetes, type II diabetes, gestational diabetes, obesity-related diabetes), hyperlipidemia (for example, hypertriglyceridemia, hypercholesterolemia, high LDL-cholesterolemia, low HDL-cholesterolemia, postprandial hyperlipidemia), hypertension, heart failure, complications of diabetes [for example, neuropathy, nephropathy, Retinopathy, diabetic cardiomyopathy, cataract, macroangiopathy, osteopenia, hyperosmotic diabetic coma, infectious diseases (e.g., respiratory tract infection, urinary tract infection, gastrointestinal infection, superficial soft tissue infection, lower limb infection), diabetic gangrene, xerostomia, hearing loss, cerebrovascular disease, peripheral blood circulation disease, metabolic syndrome (with three or more disease states selected from hypertriglyceridemia (TG), low HDL cholesterolemia (HDL-C), hypertension, abdominal obesity and impaired glucose tolerance), muscle loss, etc.

Examples of symptomatic obesity include endocrine obesity (e.g., Cushing's syndrome, hypothyroidism, insulinoma, obese type II diabetes, pseudohypoparathyroidism, hypogonadism), central obesity (e.g., hypothalamic obesity, frontal lobe syndrome, Klebsiella pneumoniae syndrome), genetic obesity (e.g., Prader-Willi syndrome, Raul-Moore-Bieder syndrome), drug-induced obesity (e.g., steroid, phenothiazine, insulin, sulfonylurea (SU) drugs, beta-blocker-induced obesity), and the like.

Examples of disease states or diseases associated with obesity include: glucose tolerance disorders, diabetes (especially type II diabetes, obese diabetes), lipid metabolism disorders (synonymous with the above-mentioned hyperlipidemia), hypertension, heart failure, hyperuricemia, fatty liver (including non-alcoholic hepatitis), coronary heart disease (myocardial infarction, angina pectoris), cerebral infarction (cerebral thrombosis, transient ischemic attack), bone/joint diseases (osteoarthritis of the knee, hip arthritis, spondylitis ankylosing, low back pain), sleep apnea syndrome/Pickwick syndrome, menstrual disorders (irregular menstrual cycles, amenorrhea, abnormal menstrual symptoms), metabolic syndrome, etc.

The compound provided by the present invention has GLP-1/GIP dual agonist activity, a long half-life, and shows a longer duration of drug effect.

The compounds of the present invention can react with any of a variety of inorganic and organic acids to form pharmaceutically acceptable acid addition salts. Pharmaceutically acceptable salts and common methods for preparing them are well known in the art.

The amino acid sequence of the present invention contains the standard single-letter or three-letter codes of twenty natural amino acids. In addition, "Aib" is α-aminoisobutyric acid, "αMePhe" is α-methylphenylalanine, "Pya(4)" is 4-pyridylalanine, "Cha" is cyclohexylalanine, and "αMeTyr" is α-methyltyrosine.

The "AEEA" mentioned in the present invention is the abbreviation of 2-(2-amino-ethoxy)-ethoxy]-acetyl. The "Ste" mentioned in the present invention represents octadecane diacyl-C(O)-C ₁₆ H ₃₂ -C(O)- derived from octadecane dioic acid.

The "effective amount" or "therapeutically effective amount" of the present invention is the amount or dosage of the compound of the present invention or a pharmaceutically acceptable salt thereof, which, when administered to a patient in a single or multiple doses, provides the desired effect in the patient being diagnosed or treated.

The "effective dose" mentioned in the present invention refers to the minimum dose that has a pharmacological effect. Specifically, in the db/db mouse multiple drug administration efficacy experiment in the present invention, it refers to the minimum dose at which the blood glucose AUC of the test substance administration group has a significant difference (P < 0.05) compared with the vehicle group.

The "Efficacy Dose" mentioned in the present invention refers to the dosage of the drug that has a pharmacological effect. Specifically, in the db/db mouse multiple drug administration efficacy experiment of the present invention, it refers to the dosage at which the blood glucose AUC of the test substance administration group has a significant difference (P < 0.05) compared with the vehicle group.

The "Vehicle group" mentioned in the present invention refers to the vehicle control group.

The "AUC" mentioned in the present invention refers to the area under the drug-time curve, that is, the area enclosed by the drug concentration curve on the time axis. This parameter is an important indicator for evaluating the degree of drug absorption and reflects the exposure characteristics of the drug in the body.

The "MRT" mentioned in the present invention refers to the mean residence time, which is the average of the time that drug molecules stay in the body, indicating the time required to eliminate 63.2% of the drug from the body.

The "MTD" mentioned in the present invention refers to the maximum tolerated dose, which is the highest dose that does not cause death in the test animals.

The "internal standard" mentioned in the present invention refers to a known amount of a pure compound added to a sample to correct errors caused by instrument signal fluctuations, human operation, etc.

The "blood glucose AUC inhibition rate" mentioned in the present invention is the percentage of blood glucose AUC reduction in the test substance administration group compared to the Vehicle group. Blood glucose AUC inhibition rate = (blood glucose AUC _Vehicle - blood glucose AUC _{test substance} ) / blood glucose AUC _Vehicle

The "titration strategy" described in the present invention is a method of dose escalation, that is, after the initial dose, the frequency of dose escalation and the dosage are adjusted to obtain the optimal dosage.

The "patient" mentioned in the present invention includes humans or animals, preferably humans.

The "modified chain" of the present invention is ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -Z, wherein each a is independently an integer from 0 to 5, each b is independently an integer from 0 to 5, and each c is independently an integer from 10 to 22; further wherein a is independently selected from an integer from 1, 2, 3, 4 or 5, b is independently selected from an integer from 1, 2, 3, 4 or 5, and c is independently selected from an integer from 12 to 22; in addition, the present invention provides a compound wherein c is 14, 16, 18 or 20, wherein Z is independently selected from -CH ₃ , carboxylic acid or carboxylic acid bioisostere, phosphate/ester or sulfonate/ester, for example, but not limited to, the Z may include carboxylic acid (-CO ₂ H) or a carboxylic acid bioisostere (for example ), phosphoric acid (-P(O)(OH) ₂ ) or sulfonic acid (-SO ₂ OH) group, preferably -CO ₂ H.

Carboxylic acid bioisosteres, suitable carboxylic acid bioisosteres are known in the art. Preferably, the bioisostere has a proton with a _pKa similar to that of the corresponding carboxylic acid. Examples of suitable bioisosteres may include, but are not limited to, tetrazoles, acylsulfonamides, acylhydroxylamides, and squaric acid derivatives, as shown below: , , , , R is Me or CF ₃ .

For the peptides mentioned herein, according to conventional peptide notation, the left end is the N-terminus (amino terminus) and the right end is the C-terminus (carboxyl terminus). The C-terminus of the peptide may be any of amide (-CONH ₂ ), carboxyl (-COOH), carboxylate (-COO ^- ), alkylamide (-CONHR') and ester (-COOR'), R' being a C _1-8 alkyl group. In particular, amide (-CONH ₂ ) is preferred.

The "coupling" or "accession" in the preparation method of the present invention refers to the process of adding a new amino acid to the bound amino acid or peptide.

Example 1: YX ₁ -EGTX ₂ -TSDYX ₃ -IX ₄ -LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein X ₁ is Aib; X ₂ is αMePhe; X ₃ is Aib; X ₄ is Aib; K at position 24 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) ₂ -(γ-Glu) ₃ -CO-(CH ₂ ) ₁₆ -CO ₂ H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 15). The above structure contains standard single-letter amino acid codes except for residues Aib2, αMePhe6, Aib11, Aib13, and K24, for which the structures have been unfolded.

The preparation method adopts the Fmoc solid phase peptide synthesis strategy, which includes:

(1) Synthesis of peptide resin intermediate 1 Take Fmoc-Rink Linker-Nle-MBHA resin (S=0.49 mmol/g), dissolve it with an appropriate amount of DCM, wash it with DCM 2-3 times, deprotect it with 20% PIP/DMF solution for 30 min, filter and wash it to obtain Fmoc-free _NH2 -Rink linker-Nle-MBHA resin, and drain the solvent for use. Take 4 equivalents of Fmoc-Lys(Boc)-OH and HOBt respectively, dissolve them in an appropriate amount of DMF/DCM; take another 4 equivalents of DIC, dilute it by one half with DCM, slowly add it to the DMF/DMC solution while stirring, and react it at -5~0℃ for not less than 60 minutes, and set it aside after activation. Add the activated Fmoc-Lys(Boc)-OH solution to NH ₂ -Rink linker–Nle-MBHA resin, control the reaction temperature at 10-30°C, couple for 240-480 min, filter and wash to obtain Fmoc-Lys(Boc)-Rink linker–Nle-MBHA resin, deprotect with 20% PIP/DMF solution for 30 min, filter and wash to obtain Lys(Boc)-Rink linker–Nle-MBHA resin without Fmoc. Under the same reaction conditions, start coupling from the second amino acid at the C-terminus to the N-terminus one by one. If the coupling is incomplete (color reaction), use HBTU/DIEA for a second condensation to ensure that each amino acid is completely condensed. The coupling order is Fmoc-Ser(tBu)-OH, Fmoc-Pro-OH, Fmoc-Pro-OH, Fmoc-Pro-OH, Fmoc-Ala-OH.H ₂ O, Fmoc-Gly-OH, Fmoc-Ser(tBu)-OH, Fmoc-Ser(tBu)-OH, Fmoc-Pro-OH, Fmoc-Gly-Gly-OH, Fmoc-Lys(Boc)-OH, Fmoc-Leu-OH, Fmoc-Leu-OH, Fmoc-Trp(Boc)-OH, Fmoc-Lys(Mtt)-OH, Fmoc-Val-OH, Fmoc-Phe-OH, Fmoc-Glu(OtBu)-OH.H ₂ O, Fmoc-Ala-OH.H ₂ O, Fmoc-Gln(Trt)-OH, Fmoc-Ala-OH.H ₂ O, Fmoc-Gln(Trt)-OH, Fmoc-Lys(Boc)-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Leu-OH, Fmoc-Aib-OH, Fmoc-Ile-OH, Fmoc-Aib-OH, Fmoc-Tyr(tBu)-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Ser(tBu)-OH, Fmoc-Thr(tBu)-OH, Fmoc-α-Me-Phe-OH, Fmoc-Thr(tBu)-OH, Fmoc-Glu(OtBu)-Gly-OH, Fmoc-Aib-OH, Boc-Tyr(tBu)-OH. The following was obtained: Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-αMePhe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Aib-Ile-Aib-Leu-Asp(OtBu)-Lys(Boc)-Gln(Trt)-Ala-Gln(Trt)-Ala-Glu(OtBu)-Phe-Val-Lys(Mtt)-Trp(Boc)-Leu-Leu-Lys(Boc)-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Lys(Boc)-Rink Linker-Nle-MBHA Resin After the above condensation was completed, 50% HFIP/DCM solution was used to remove the Mtt protection for 30 minutes, followed by washing and filtration to obtain peptide resin intermediate 1: Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-αMePhe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Aib-Ile-Aib-Leu-Asp(OtBu)-Lys(Boc)-Gln(Trt)-Ala-Gln(Trt)-Ala-Glu(OtBu)-Phe-Val- Lys-Trp(Boc)-Leu-Leu-Lys(Boc)-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Lys(Boc)-Rink Linker-Nle-MBHA Resin.

(2) Modification Chain Modification Step Take 4 equivalents of Fmoc-AEEA-OH and HOBt and dissolve them in an appropriate amount of DMF/DCM; take another 4 equivalents of DIC, dilute it by one half with DCM, slowly add it to the DMF/DCM solution while stirring, and stir the reaction at -5 to 0°C for no less than 60 minutes, and set aside after activation. Add the activated Fmoc-AEEA-OH solution to the previously dissolved and washed peptide resin intermediate 1, control the reaction temperature at 10 to 30°C, and couple the reaction for 240 to 480 minutes, filter and wash, remove the Fmoc protection with 20% PIP/DMF solution for 30 minutes, and filter and wash. According to the above reaction conditions, the activated Fmoc-AEEA-OH, Fmoc-AEEA-OH, Fmoc-Glu(α-OtBu)-OH, Fmoc-Glu(α-OtBu)-OH, Fmoc-Glu(α-OtBu)-OH and octadecane dioic acid mono-tert-butyl ester were coupled on the resin in sequence, Fmoc was deprotected, and the mixture was washed with DCM and dried to obtain P015 peptide resin: Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-αMePhe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Aib-Ile-Aib-Leu-Asp(OtBu)-Lys(Boc)-Gln(Trt)-Ala-Gln(Trt)-Ala-Glu(OtBu)-Phe-Val-Lys(tBuO-Ste -γ-Glu(α-OtBu)-γ-Glu(α-OtBu)-γ-Glu(α-OtBu)-AEEA-AEEA)-Trp(Boc)-Leu-Leu-Lys(Boc)-Gly-Gly-Pro- Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Lys(Boc)-Rink-Linker-Nle-MBHA Resin. Take P 015 peptide resin, add 12-15 mL/g peptide resin lysis agent (TFA:EDT:TIS:H ₂ O, volume ratio 94:2:2:2), stir and react at 25±5℃ for 4 hours, filter the reaction mixture using a sand core funnel, collect the filtrate, wash the resin with a small amount of TFA 3 times, combine the filtrate and reduce the pressure to concentrate, add methyl tert-butyl ether (MBTE) for precipitation, and then wash with MBTE 3-4 times, evaporate MTBE, and reduce the pressure to dry the crude product at room temperature to constant weight to obtain P015 crude product. After subsequent purification, dry to obtain P015 sample (mass spectrum MS: 5255.2).

Example 2: YX ₁ -EGTX ₂ -TSDYX ₃ -IX ₄ -LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein X ₁ is Aib; X ₂ is αMePhe; X ₃ is Aib; X ₄ is Aib; K at position 28 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) ₃ -(γ-Glu) ₁ -CO-(CH ₂ ) ₁₆ -CO ₂ H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 19). The above structure contains standard single-letter amino acid codes except for residues Aib2, αMePhe6, Aib11, Aib13 and K28, for which the structures have been unfolded.

The preparation method adopts the Fmoc solid phase peptide synthesis strategy, which includes:

(1) Synthesis of peptide resin intermediate 2 Take Fmoc-Rink Linker-Nle-MBHA resin (S=0.49 mmol/g), dissolve it with an appropriate amount of DCM, wash it with DCM 2-3 times, deprotect it with 20% PIP/DMF solution for 30 min, wash it and filter it to obtain the Fmoc-free _NH2 -Rink linker-Nle-MBHA resin, and drain the solvent for later use. Take 4 equivalents of Fmoc-Lys(Boc)-OH and HOBt respectively, dissolve them in an appropriate amount of DMF/DCM; take another 4 equivalents of DIC, dilute it by one half with DCM, slowly add it to the DMF/DMC solution while stirring, stir it at -5~0℃ for not less than 60 minutes, and set it aside after activation. Add the activated Fmoc-Lys(Boc)-OH solution to NH ₂ -Rink linker–Nle-MBHA resin, control the reaction temperature at 10-30°C, couple for 240-480 min, filter and wash to obtain Fmoc-Lys(Boc)-Rink linker–Nle-MBHA resin, deprotect with 20% PIP/DMF solution for 30 min, filter and wash to obtain Lys(Boc)-Rink linker–Nle-MBHA resin without Fmoc. Under the same reaction conditions, start coupling from the second amino acid at the C-terminus to the N-terminus one by one. If the coupling is incomplete (color reaction), use HBTU/DIEA for a second condensation to ensure that each amino acid is completely condensed. The coupling order is Fmoc-Ser(tBu)-OH, Fmoc-Pro-OH, Fmoc-Pro-OH, Fmoc-Pro-OH, Fmoc-Ala-OH.H ₂ O, Fmoc-Gly-OH, Fmoc-Ser(tBu)-OH, Fmoc-Ser(tBu)-OH, Fmoc-Pro-OH, Fmoc-Gly-Gly-OH, Fmoc-Lys(Mtt)-OH, Fmoc-Leu-OH, Fmoc-Leu-OH, Fmoc-Trp(Boc)-OH, Fmoc-Lys(Boc)-OH, Fmoc-Val-OH, Fmoc-Phe-OH, Fmoc-Glu(OtBu)-OH.H ₂ O, Fmoc-Ala-OH.H ₂ O, Fmoc-Gln(Trt)-OH, Fmoc-Ala-OH.H ₂ O, Fmoc-Gln(Trt)-OH, Fmoc-Lys(Boc)-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Leu-OH, Fmoc-Aib-OH, Fmoc-Ile-OH, Fmoc-Aib-OH, Fmoc-Tyr(tBu)-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Ser(tBu)-OH, Fmoc-Thr(tBu)-OH, Fmoc-α-Me-Phe-OH, Fmoc-Thr(tBu)-OH, Fmoc-Glu(OtBu)-Gly-OH, Fmoc-Aib-OH, Boc-Tyr(tBu)-OH. The following was obtained: Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-αMePhe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Aib-Ile-Aib-Leu-Asp(OtBu)-Lys(Boc)-Gln(Trt)-Ala-Gln(Trt)-Ala-Glu(OtBu)-Phe-Val-Lys(Boc)-Trp(Boc)-Leu-Leu-Lys(Mtt)-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Lys(Boc)-Rink Linker-Nle-MBHA Resin After the above condensation was completed, 50% HFIP/DCM solution was used to remove the Mtt protection for 30 minutes, followed by washing and filtration to obtain peptide resin intermediate 2: Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-αMePhe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Aib-Ile-Aib-Leu-Asp(OtBu)-Lys(Boc)-Gln(Trt)-Ala-Gln(Trt)-Ala-Glu(OtBu)-Phe-Val-Lys(Boc)-Trp(Boc)-Leu-Leu- Lys-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Lys(Boc)-Rink Linker-Nle-MBHA Resin.

(2) Modification Chain Modification Step Take 4 equivalents of Fmoc-AEEA-OH and HOBt and dissolve them in an appropriate amount of DMF/DCM; take another 4 equivalents of DIC, dilute it by one half with DCM, slowly add it to the DMF/DCM solution while stirring, and stir the reaction at -5 to 0°C for no less than 60 minutes, and set aside after activation. Add the activated Fmoc-AEEA-OH solution to the previously dissolved and washed peptide resin intermediate 2, control the reaction temperature at 10 to 30°C, and couple the reaction for 240 to 480 minutes, filter and wash, remove the Fmoc protection with 20% PIP/DMF solution for 30 minutes, and filter and wash. According to the above reaction conditions, the activated Fmoc-AEEA-OH, Fmoc-AEEA-OH, Fmoc-AEEA-OH, Fmoc-Glu(α-OtBu)-OH and octadecane dioic acid mono-tert-butyl ester were coupled on the resin, Fmoc was deprotected, and the mixture was washed with DCM and dried to obtain the P019 peptide resin: Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-αMePhe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Aib-Ile-Aib-Leu-Asp(OtBu)-Lys(Boc)-Gln(Trt)-Ala-Gln(Trt)-Ala-Glu(OtBu)-Phe-Val-Lys(tBuO-Ste -γ-Glu(α-OtBu)-AEEA-AEEA-AEEA)-Trp(Boc)-Leu-Leu-Lys(Boc)-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Lys(Boc)-Rink-Linker-Nle-MBHA Resin. Take P 019 peptide resin, add 12-15 mL/g peptide resin lysis agent (TFA:EDT:TIS:H ₂ O, volume ratio 94:2:2:2), stir and react at 25±5℃ for 4 hours, filter the reaction mixture using a sand core funnel, collect the filtrate, wash the resin with a small amount of TFA 3 times, combine the filtrate and reduce the pressure to concentrate, add methyl tert-butyl ether (MBTE) for precipitation, and then wash with MBTE 3-4 times, evaporate MTBE, and reduce the pressure to dry the crude product at room temperature to constant weight to obtain P019 crude product. After subsequent purification, dry to obtain P019 sample (mass spectrum MS: 5142.0).

Example 3 YX ₁ -EGTX ₂ -TSDYX ₃ -IX ₄ -LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein X ₁ is Aib; X ₂ is αMePhe; X ₃ is Aib; X ₄ is Aib; K at position 24 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) ₂ -(γ-Glu) ₁ -CO-(CH ₂ ) ₁₆ -CO ₂ H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 7). The above structure contains standard single-letter amino acid codes except for residues Aib2, αMePhe6, Aib11, Aib13, and K24, for which the structures have been unfolded.

Similar to the above Example 1, the peptide resin intermediate 1 was used to carry out the preparation steps to synthesize the peptide of SEQ ID NO: 7 of the present invention (mass spectrum MS: 4997.2). The difference is that the modification process of the modified chain is to sequentially couple Fmoc-AEEA-OH, Fmoc-AEEA-OH, Fmoc-Glu (α-OtBu) -OH and octadecane dioic acid mono-tert-butyl ester on the resin, deprotect Fmoc, wash with DCM and dry to obtain P007 peptide resin, and the other steps are similar.

Example 4: _YX1 - _EGTX2 -TSDYX3- _{IX4 -} LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at position 24 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _2- (γ-Glu) ₁ -CO-( _CH2 ) ₁₈ - _CO2H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 13). The above structure contains standard single-letter amino acid codes except for residues Aib2, αMePhe6, Aib11, Aib13, and K24, for which the structures have been unfolded.

Similar to the above Example 1, the peptide resin intermediate 1 was used to carry out the preparation steps to synthesize the peptide of SEQ ID NO: 13 of the present invention (mass spectrum MS: 5025.2). The difference is that the modification process of the modified chain is to sequentially couple Fmoc-AEEA-OH, Fmoc-AEEA-OH, Fmoc-Glu (α-OtBu) -OH and mono-tert-butyl eicosanedioate on the resin, deprotect Fmoc, wash with DCM and dry to obtain P013 peptide resin, and the other steps are similar.

Example 5: YX ₁ -EGTX ₂ -TSDYX ₃ -IX ₄ -LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein X ₁ is Aib; X ₂ is αMePhe; X ₃ is Aib; X ₄ is Aib; K at position 24 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) ₂ -(γ-Glu) ₁ -CO-(CH ₂ ) ₂₀ -CO ₂ H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 14). The above structure contains standard single-letter amino acid codes except for residues Aib2, αMePhe6, Aib11, Aib13, and K24, for which the structures have been unfolded.

Similar to the above Example 1, peptide resin intermediate 1 was used to carry out the preparation steps to synthesize the peptide of SEQ ID NO: 14 of the present invention (mass spectrum MS: 5052.4). The difference is that the modification process of the modified chain is to sequentially couple Fmoc-AEEA-OH, Fmoc-AEEA-OH, Fmoc-Glu (α-OtBu)-OH and docosane dioic acid mono-tert-butyl ester on the resin, deprotect Fmoc, wash with DCM and dry to obtain P014 peptide resin, and the other steps are similar.

Example 6: YX ₁ -EGTX ₂ -TSDYX ₃ -IX ₄ -LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein X ₁ is Aib; X ₂ is αMePhe; X ₃ is Aib; X ₄ is Aib; K at position 24 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) ₃ -(γ-Glu) ₁ -CO-(CH ₂ ) ₂₀ -CO ₂ H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 16). The above structure contains standard single-letter amino acid codes except for residues Aib2, αMePhe6, Aib11, Aib13, and K24, for which the structures have been unfolded.

Similar to the above Example 1, the peptide resin intermediate 1 was used to carry out the preparation steps to synthesize the peptide of SEQ ID NO: 16 of the present invention (mass spectrum MS: 5198.0). The difference is that the modification process of the modified chain is to sequentially couple Fmoc-AEEA-OH, Fmoc-AEEA-OH, Fmoc-AEEA-OH, Fmoc-Glu (α-OtBu)-OH and docosanediolatoic acid mono-tert-butyl ester on the resin, deprotect Fmoc, wash with DCM and dry to obtain P016 peptide resin, and the other steps are similar.

Example 7: _YX1 - _EGTX2 -TSDYX3- _{IX4 -} LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at position 28 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _2- (γ-Glu) ₁ -CO-( _CH2 ) ₁₆ - _CO2H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO:8). The above structure contains standard single-letter amino acid codes except for residues Aib2, αMePhe6, Aib11, Aib13, and K28, for which the structures have been unfolded.

Similar to the above Example 2, peptide resin intermediate 2 was used to carry out the preparation steps to synthesize the peptide of SEQ ID NO: 8 of the present invention (mass spectrum MS: 4996.8). The difference is that the modification process of the modified chain is to sequentially couple Fmoc-AEEA-OH, Fmoc-AEEA-OH, Fmoc-Glu (α-OtBu) -OH and octadecane dioic acid mono-tert-butyl ester on the resin, deprotect Fmoc, wash with DCM and dry to obtain P008 peptide resin, and the other steps are similar.

Example 8: _YX1 - _EGTX2 -TSDYX3- _{IX4 -} LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at position 28 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _2- (γ-Glu) ₁ -CO-( _CH2 ) ₂₀ - _CO2H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 17). The above structure contains standard single-letter amino acid codes except for residues Aib2, αMePhe6, Aib11, Aib13 and K28, for which the structures have been unfolded.

Similar to the above Example 2, peptide resin intermediate 2 was used to carry out the preparation steps to synthesize the peptide of SEQ ID NO: 17 of the present invention (mass spectrum MS: 5052.8). The difference is that the modification process of the modified chain is to sequentially couple Fmoc-AEEA-OH, Fmoc-AEEA-OH, Fmoc-Glu (α-OtBu) -OH and docosane diacid mono-tert-butyl ester on the resin, deprotect Fmoc, wash with DCM and dry to obtain P017 peptide resin, and the other steps are similar.

Example 9: _YX1 - _EGTX2 - _TSDYX3 - _IX4- LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at position 28 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _2- (γ-Glu) ₃ -CO-( _CH2 ) ₁₆ - _CO2H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 18). The above structure contains standard single-letter amino acid codes except for residues Aib2, αMePhe6, Aib11, Aib13, and K28, for which the structures have been unfolded.

Similar to the above Example 2, peptide resin intermediate 2 was used to carry out the preparation steps to synthesize the peptide of SEQ ID NO: 18 of the present invention (mass spectrum MS: 5255.2). The difference is that the modification process of the modified chain is to sequentially couple Fmoc-AEEA-OH, Fmoc-AEEA-OH, Fmoc-Glu(α-OtBu)-OH, Fmoc-Glu(α-OtBu)-OH, Fmoc-Glu(α-OtBu)-OH and octadecane dioic acid mono-tert-butyl ester on the resin, deprotect Fmoc, wash with DCM and dry to obtain P018 peptide resin, and the other steps are similar.

Example 10: _YX1 - _EGTX2 -TSDYX3- _{IX4 -} LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at position 28 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _3- (γ-Glu) ₁ -CO-( _CH2 ) ₂₀ - _CO2H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO:20). The above structure contains standard single-letter amino acid codes except for residues Aib2, αMePhe6, Aib11, Aib13 and K28, for which the structures have been unfolded.

Similar to the above Example 2, peptide resin intermediate 2 was used to carry out the preparation steps to synthesize the peptide of SEQ ID NO: 20 of the present invention (mass spectrum MS: 5198.4). The difference is that the modification process of the modified chain is to sequentially couple Fmoc-AEEA-OH, Fmoc-AEEA-OH, Fmoc-AEEA-OH, Fmoc-Glu (α-OtBu)-OH and docosane dioic acid mono-tert-butyl ester on the resin, deprotect Fmoc, wash with DCM and dry to obtain P020 peptide resin, and the other steps are similar.

In some embodiments of the present invention, peptide resin intermediate 3 is prepared by referring to the preparation method of peptide resin intermediate 1 in Example 1 to obtain peptide resin intermediate 3 (peptide resin intermediate with a modified chain at position 16 Lys): Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-αMePhe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Aib-Ile-Aib-Leu-Asp(OtBu)-Lys-Gln(Trt)-Ala-Gln(Trt)-Ala-Glu(OtBu)-Phe-Val-Lys(Boc)-Trp(Boc)-Leu-Leu-Lys(Boc)-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Lys(Boc)-Rink Linker-Nle-MBHA Resin Peptide resin intermediate 4, refer to the preparation method of peptide resin intermediate 1 in Example 1 to obtain peptide resin intermediate 4 (peptide resin intermediate with a modified chain at position 40 Lys): Boc-Tyr(tBu)-Aib-Glu(OtBu)-Gly-Thr(tBu)-αMePhe-Thr(tBu)-Ser(tBu)-Asp(OtBu)-Tyr(tBu)-Aib-Ile-Aib-Leu-Asp(OtBu)-Lys(Boc)-Gln(Trt)-Ala-Gln(Trt)-Ala-Glu(OtBu)-Phe-Val-Lys(Boc)-Trp(Boc)-Leu-Leu-Lys(Boc)-Gly-Gly-Pro-Ser(tBu)-Ser(tBu)-Gly-Ala-Pro-Pro-Pro-Ser(tBu)-Lys-Rink Linker-Nle-MBHA Resin Other dual agonist compounds in the present invention can be prepared by referring to the above method.

Related Assays The conditions and data used in the Examples for several assays are provided below.

1. Extracorporeal Function

(I) In vitro binding activity to human GLP-1 and GIP receptors The in vitro binding potency of the compounds of the present invention to human GIP and GLP-1 receptors was evaluated by measuring the binding affinity Ki using crude cell membranes obtained from cloned cell lines expressing human GLP-1R cDNA or human GIP-R cDNA. 1) In vitro binding activity to human GLP-1 receptor hGLP-1 and the compounds of the present invention were dissolved in DMSO and stored at -80°C. 89 μL of the membrane (5 μg/well) dissolved in binding buffer (50 mM Hepes, pH 7.4, 5 mM MgCl ₂ , 5 mM EDTA, 0.005% TWEEN, 0.005% HSA) was transferred to a 96-well assay plate. Compounds were serially diluted in DMSO, and 1 μL of the diluted compound or 100% DMSO was added to the assay plate containing the membrane solution. 10 μL of [ ^125I ]GLP-1 was then added (final reaction concentration was 0.15 nM). The assay plate was placed at room temperature for 90 minutes. The membrane complexes were harvested using Cell Harvester onto 0.5% PEI pre-coated GF/B plates and washed three times with 500 μL of 4°C pre-cooled wash buffer (50 mM Hepes, pH 7.4, 500 mM NaCl). After drying at 37°C for 2 hours, 50 μL of scintillation buffer was added to each well, sealed and left for at least 1 hour. The membrane-bound radioligand level was then determined using Microbet2. The absolute IC ₅₀ concentration was obtained by nonlinear regression of the percentage of [ ¹²⁵ I]GLP-1 binding and the concentration of the added compound. The IC ₅₀ concentration was converted to Ki (Ki is the inhibition constant) using the Cheng-Prusoff formula. 2) In vitro binding activity to human GIP receptor hGIP and the compounds of the present invention were dissolved in DMSO and stored at -80°C. 98 μL of the membrane (15 μg/well) dissolved in binding buffer (50 mM HEPES pH 7.4, 5 mM MgCl ₂ , 1 mM CaCl ₂ , 0.1% BSA, 0.005% Tween-20) was transferred to a 96-well assay plate. Compounds were serially diluted in DMSO, and 2 μL of the diluted compound or 100% DMSO was added to the assay plate containing the membrane solution. 100 μL of [ ¹²⁵ I]GIP was then added (final reaction concentration was 0.0315 nM). The assay plate was incubated at room temperature for 90 min. The membrane complexes were harvested using a Cell Harvester onto 0.5% PEI pre-coated GF/B plates and washed three times with 500 μL of 4°C pre-cooled wash buffer (50 mM Tris-HCl pH 7.4, 125 mM NaCl). After drying at 37°C for 2 h, 50 μL of scintillation buffer was added to each well, sealed, and left for at least 1 h. The membrane-bound radioligand level was then determined by reading with a Microbeta2. Absolute IC ₅₀ concentrations were obtained by nonlinear regression of the percentage of [ ¹²⁵ I]GIP bound and the concentration of the added compound. IC ₅₀ concentrations were converted to Ki (Ki is the inhibition constant) using the Cheng-Prusoff equation. Table 1. Receptor binding affinity, Ki ratio Compound GLP-1R GIPR Ki Ki GLP-1 1.00 NA GIP NA 1.00 P001 19.52 8.82 Tirzepatide 0.28 0.06 P007 0.47 NA P008 1.09 0.16 P013 0.28 0.14 P014 0.37 0.08 P015 0.17 NA P016 0.63 0.11 P017 0.56 0.04 P018 1.45 0.05 P019 0.93 0.09 P020 1.36 0.05 NA: Not Detected Ki ratio: The ratio of the Ki value of the endogenous ligand to the Ki value of the test compound. The binding affinity of all compounds to the receptor is expressed as Ki ratio. The larger the Ki ratio, the stronger the binding affinity of the compound. Compared with P001, the binding affinity of the compounds of the present invention to human GLP-1R and GIPR has decreased to a certain extent (see Table 1). Compared with Tirzepatide, except for P015, the binding affinity of the other 9 compounds of the present invention to human GLP-1R is slightly stronger than that of Tirzepatide, and the binding affinity of the five compounds P008, P013, P014, P016 and P019 to GIPR is also slightly stronger than that of Tirzepatide (see Table 1). In summary, the binding affinity of the compounds of the present invention with different forms of modified chains to human GLP-1R and GIPR is somewhat lower than that of P001, but slightly stronger than or equivalent to that of Tirzepatide.

(II) Stimulating activity on hGLP-1R and hGIPR For human GLP-1 and GIP receptors, the in vitro functional activity of the compounds of the present invention on these receptors was determined in HEK-293 clone cell lines expressing these receptors. 1) Stimulating activity on human GLP-1 receptor (cAMP reporter gene method) The stimulating activity of the compounds of the present invention on human GLP-1R was determined using the cAMP reporter gene method, and P001, Tirzepatide and endogenous ligand GLP-1 were used as references. HEK293/CRE/GLP-1R cells were inoculated in a 96-well plate at 50,000 cells/well (80 μL/well) and cultured overnight in a 37°C, 5% CO ₂ incubator. Add 20 μL/well of the test medium (DMEM containing 0.1% casein) containing the compound (compound of the present invention, P001, Tirzepatide or GLP-1) to the 96-well plate, continue to incubate in a CO ₂ incubator at 37°C for 6 hours, equilibrate to room temperature, remove the supernatant, add 50 μL/well Bright-Glo reagent, and shake and lyse at room temperature for 10 minutes. Use Envision enzyme labeler to read luminescence and measure luciferase activity. The response value of 100 nM GLP-1 is set as 100% response value, and GraphPad is used to perform nonlinear regression based on the response percentage and the concentration of the added compound to obtain the EC ₅₀ value of each compound. 2) Stimulating activity on human GIP receptor (LANCE Ultra cAMP assay) The LANCE Ultra cAMP Kit was used to measure the stimulating activity of the compounds of the present invention on human GIPR, and P001, Tirzepatide and endogenous ligand GIP were used as references. The in vitro stimulating activity of the compounds of the present invention on GIPR was measured in HEK293 cells stably expressing human GIPR (HEK293/GIPR cells). HEK293/GIPR cells were prepared with HBSS buffer (0.1% Casein, 500 μM IBMX, 5 mM HEPES) and inoculated in a 384-well cell culture plate at 1000 cells/well (5 μL/well). 5 μL of HBSS buffer containing 2× compound was added to the above 384-well cell culture plate. After sealing the plate, place it in a 5% CO ₂ incubator at 37°C for about 30 minutes. After the incubation is completed, add 5 μL of cAMP-Eu working solution and 5 μL of cAMP-Ulight working solution in turn, and shake the plate to mix. Incubate at 25°C for 1 hour, read the signal values at 665 nm and 615 nm on the Envision enzyme marker, calculate the 665 nm/615 nm ratio, and convert it to cAMP concentration using the cAMP standard curve. The response value of 1 μM GIP was set to 100% response value, and GraphPad was used to perform nonlinear regression based on the response percentage and the added compound concentration to obtain the EC ₅₀ value of each compound. Table 2. Stimulating activity on human GLP-1 and GIP receptors, EC ₅₀ values Compound GLP-1R GIPR _EC50 value _EC50 value GLP-1 1.00 NA GIP NA 1.00 P001 52.09 7.13 Tirzepatide 0.74 0.51 P007 5.84 1.15 P008 4.79 1.67 P013 2.41 0.86 P014 1.73 0.25 P015 2.44 0.17 P016 1.56 0.56 P017 1.35 0.16 P018 3.63 2.55 P019 4.95 1.68 P020 1.41 0.22 NA: Not Detected EC ₅₀ Value: Ratio of the EC ₅₀ value of the endogenous ligand to the EC ₅₀ value of the test compound The stimulatory activity of all compounds on human GLP-1 and GIP receptors is expressed as EC ₅₀ value, and the larger the EC ₅₀ value, the stronger the activity of the compound. Compared with P001, the stimulatory activity of the compounds of the present invention on human GLP-1R and GIPR has decreased to a certain extent (see Table 2). Compared with Tirzepatide, the stimulatory activity of all compounds of the present invention on human GLP-1R is stronger than that of Tirzepatide, and the stimulatory activity of the six compounds P007, P008, P013, P016, P018 and P019 on human GIPR is also slightly stronger than that of Tirzepatide (see Table 2). In summary, the compounds of the present invention with different forms of chain modification have different degrees of lower stimulatory activity on human GLP-1R and GIPR compared with P001, but 6 compounds are stronger than Tirzepatide. In vitro functional tests show that the activities of the 10 compounds of the present invention are not much different, so the single-dose hypoglycemic experiment with db/db mice was used to further screen the compounds.

2. Pharmacokinetics

(I) Pharmacokinetics of single-dose administration in SD rats Plasma was collected and the plasma drug concentration was tested after subcutaneous or intravenous administration to elucidate the pharmacokinetic properties of the compounds of the present invention in vivo. The compounds were dissolved in phosphate buffer and filtered through a filter membrane (PTFE, 0.45 μm) to obtain a 25 nmol/mL compound solution. Male SD rats (230-268 g, n=3) were given 50.0 nmol/kg subcutaneously (sc) or 25.0 nmol/kg intravenously (iv). Approximately 150 μL of whole blood was collected from the cervical vein in EDTA-K ₂ anticoagulant tubes at predose, 0.0833 (iv), 0.25, 0.5, 1, 2, 4, 6, 8, 24, 48, 72, 96, and 120 h. Blood samples were centrifuged at 1500 g for 10 min to obtain plasma, which was stored at -90 ^o C to -60°C for analysis. Sample treatment (1) Sample treatment method for Tirzepatide, P007, P008, P013, and P014 Take 30.0 μL of thawed plasma sample, add 150 μL of acetonitrile solution (containing 5 ng·mL ^-1 verapamil, 50 ng·mL ^-1 glibenclamide, 200 ng·mL ^-1 tolbutamide, and 200 ng·mL ^-1 diclofenac) to precipitate protein, vortex the mixture for 5 min, and centrifuge at 3700 rpm for 8 min. Take 70.0 μL of supernatant, add 70.0 μL of 0.5% formic acid aqueous solution, vortex for 5 min, take 15 μL of the mixture, and inject it into LC-MS/MS to detect plasma drug concentration. (2) Sample processing method for P015, P016, P017, and P018: Take 20.0 μL of thawed plasma samples, add 60.0 μL of acetonitrile solution (containing 5 ng·mL ^-1 verapamil, 50 ng·mL ^-1 glibenclamide, 200 ng·mL ^-1 tolbutamide, and 200 ng·mL ^-1 diclofenac) to precipitate protein, vortex the mixture for 1 min, and centrifuge at 13000 rpm for 8 min. Take 60.0 μL of the supernatant, add 60.0 μL of 0.5% formic acid aqueous solution, vortex for 10 min, take 10.0 μL of the mixture, and inject it into LC-MS/MS to detect the plasma drug concentration. (3) Sample processing method for P019 and P020: Take 30.0 μL of thawed plasma samples, add 150 μL of acetonitrile solution (containing 5 ng·mL ^-1 verapamil, 50 ng·mL ^-1 glibenclamide, 200 ng·mL ^-1 tolbutamide and 200 ng·mL ^-1 diclofenac) to precipitate protein, vortex the mixture for 5 min, and centrifuge at 3700 rpm for 8 min. Take 70.0 μL of supernatant, add 70.0 μL of 0.5% formic acid aqueous solution, vortex for 5 min, take 10.0 μL of the mixture, and inject LC-MS/MS to detect plasma drug concentration. According to the blood drug concentration of the compound in SD rats, the pharmacokinetic parameters were calculated using the software Winnolin 8.2 with a non-compartmental model. The results are shown in Tables 3 and 4. Table 3 Pharmacokinetic parameters of different peptide compounds after subcutaneous injection in rats PK parameters Unit Tirzepatide P007 P008 P013 P014 Dosage nmol/kg 50 50 50 50 50 T1/2 h 11.4±1.79 15.0±2.89 18.7±4.82 15.0±0.737 14.8±0.503 Tmax h 7.33±1.15 7.33±1.15 13.3±9.24 24.0±0 24.0±0 Cmax nM 128±42.8 169±14.7 149±20.5 142±28.2 170±16.4 AUC0-t nM·h 3610±1200 6150±776 5400±1030 5740±1070 6230±651 AUC0-inf nM·h 3670±1190 6630±439 6680 6080±1180 6620±712 MRT0-t h 19.0±0.265 21.4±1.87 21.0±1.85 26.2±0.700 26.2±0.781 MRT0-inf h 20.3±0.755 25.7±1.69 25.4 29.9±1.33 30.0±1.01 Table 4 Pharmacokinetic parameters of different peptide compounds after subcutaneous injection in rats PK parameters Unit P015 P016 P017 P018 P019 P020 Dosage nmol/kg 50 50 50 50 50 50 T1/2 h 16.1±1.21 14.1±0.153 18.7±2.69 18.3±6.77 10.9±1.92 17.9±0.57 Tmax h 24.0±0 24.0±0 24.0±0 24.0±0 13.3±9.24 24.0±0 Cmax nM 213±21.3 119±7.00 112±15.5 175±45.1 302±89.9 139±28.1 AUC0-t nM·h 8830±862 4840±270 4190±242 7490±1360 7910±545 5620±1690 AUC0-inf nM·h 9490±1090 5070±262 4530±28.28 7940±1230 8630±601 7070±28.3 MRT0-t h 26.8±0.557 28.1±1.12 27.9±2.99 28.5±4.45 17.1±2.55 29.5±4.68 MRT0-inf h 31.4±1.35 31.1±1.00 36.9±3.46 33.4±7.00 18.3±2.76 37.1±0.636 The PK test of subcutaneous injection of the compounds of the present invention in rats showed that the half-life of P007, P008, P013, P014, P015, P016, P017, P018, and P020 was longer than that of Tirzepatide (increased by about 23.7% to 64.0%). The compounds of the present invention modified with modified chains can achieve a longer plasma half-life than Tirzepatide, a dual-target product on the market. In addition, compared with P001, the half-life of the compounds of the present invention is significantly extended, increasing by more than 15 times, more than 20 times, and even more than 30 times.

(II) Pharmacokinetics in C57 mice Plasma was collected and the blood drug concentration was measured after subcutaneous or intravenous administration of the drug to elucidate the pharmacokinetic properties of the compounds of the present invention in vivo. The compound was dissolved in phosphate buffer containing 0.1% Tween 20 and filtered through a filter membrane (PTFE, 0.45 μm) to obtain a 15 nmol/mL compound solution. C57 mice (18-22 g) were given a dose of 30 nmol/kg subcutaneously (sc) or 15.0 nmol/kg intravenously (iv). Approximately 100 μL of whole blood was collected through the eye socket in an EDTA-K ₂ anticoagulant tube at predose, 0.5, 1, 2, 4, 8, 24, 48 and 72 h. Blood samples were centrifuged at 1500 g for 10 min to obtain plasma, and the plasma was stored at -90 ^o C to -60 °C for analysis. Sample treatment (1) Sample treatment method for Tirzepatide, P014, P016, and P020 Take 20.0 μL of thawed plasma samples, add 20 μL of 50% methanol aqueous solution (containing 100 ng·mL ^-1 internal standard) and 60 μL of acetonitrile to precipitate protein, vortex the mixture for 5 min, and centrifuge at 3700 rpm for 8 min. Take 50.0 μL of the supernatant, add 50.0 μL of 0.5% formic acid aqueous solution, vortex for 5 min, take 50 μL of the mixture, and inject LC-MS/MS to detect the plasma drug concentration. According to the blood concentration of the compound in C57 mice, the pharmacokinetic parameters were calculated using the software Winnolin 8.2 with a non-compartmental model. The results are shown in Tables 5 and 6. Table 5 Average pharmacokinetic parameters after a single subcutaneous administration in mice PK parameters Unit Tirzepatide P014 P016 P020 Cmax ng/mL 871 851.84 433.67 835.67 _Tmax hr 6 6 8 4.25 t _1/2 hr 9.21 20.33 23.05 19.89 AUC _(0-t) ng·hr/mL 19413.9 27316.4 15914.8 18950.56 AUC _{(0- ∞ )} ng·hr/mL 19704.7 30150.5 18160.6 21093.26 MRT _(0-t) hr 14.48 22.94 25.98 21.84 MRT _{(0- ∞ )} hr 15.29 30.28 35.91 29.36 Table 6 Average pharmacokinetic parameters after single intravenous administration in mice PK parameters Unit Tirzepatide P014 P016 P020 C0 ng/mL 1901.94 1943.93 1147.89 1819.74 t _1/2 hr 10.47 17.20 20.15 16.66 AUC _(0-t) ng·hr/mL 12723.1 17458.9 11082.1 16281.7 AUC _{(0- ∞ )} ng·hr/mL 12822.1 18245.6 12083.3 17369.9 MRT _(0-t) hr 11.39 15.72 19.92 16.80 MRT _{(0- ∞ )} hr 12.00 19.215 26.64 21.77 The PK test of the compounds of the present invention injected subcutaneously in mice showed that the half-life of P014, P016 and P020 was longer than that of Tirzepatide (increased by about 59.2% to 92.5%), and was significantly longer than that of Tirzepatide. The compounds of the present invention modified with modified chains can achieve a longer plasma half-life than Tirzepatide, a dual-target product on the market.

(III) Pharmacokinetics in cynomolgus monkeys Plasma was collected after subcutaneous or intravenous administration of the drug to cynomolgus monkeys and its blood drug concentration was detected to clarify the pharmacokinetic properties of the compounds of the present invention in vivo. The compounds were dissolved in 1.4% PG in 8 mM Na2HPO4 Buffer (pH 7.39) and filtered through a filter membrane (PTFE, 0.45μm) to obtain 25 and 12.5 nmol/mL compound solutions, respectively. Cynomol/kg was administered subcutaneously (sc) at 25 nmol/kg or intravenously (iv) at 12.5 nmol/kg. Blood was collected from the cephalic vein or other appropriate vein at predose, 2h, 12h, 24h, 32h, 48h, 3d (72h), 4d (96h), 6d (144h), 9d (216h), 12d (288h), 15d (360h) and 18d (432h). About 500 μL of whole blood was collected in EDTA-K ₂ anticoagulant tubes. The blood samples were centrifuged at 2200 g for 10 min to obtain plasma, which was stored at -90 ^o C to -60 °C for analysis. Sample processing (1) Sample processing method for Tirzepatide, P014, P016, P017, and P020: Take 70.0 μL of thawed plasma samples, add 70 μL of acetonitrile (containing 1 ng/ml internal standard) to precipitate protein, vortex the mixture for 1 min, and centrifuge at 14,000 rpm for 10 min. Take 60.0 μL of the supernatant, add 60.0 μL of 0.5% formic acid aqueous solution, vortex for 5 min, take 10 μL of the mixture, and inject LC-MS/MS to detect plasma drug concentration. Based on the blood drug concentration of the compound in cynomolgus monkeys, the pharmacokinetic parameters were calculated using the non-compartmental model using Winnolin 8.2 software. The results are shown in Tables 7 and 8 below. Table 7 Average pharmacokinetic parameters after single subcutaneous administration in cynomolgus monkeys PK parameters Unit Tirzepatide P014 P016 P017 P020 _Tmax hr 22.67±10.10 16.00±6.93 40.00±13.90 48.00±24.00 22.70±10.10 C _max ng/mL 1297.45± 300.00 951.00±283 2997.00± 481.00 1655.00± 295.00 1349.00±77.00 T1/2 hr 62.54±6.07 52.70±6.45 60.40±12.80 44.30±4.80 67.40±9.47 AUC _(0-t) hr*ng/mL 148063.24± 38207.00 87690.00± 24556.00 319272.00± 41475.00 153476.00± 28919.00 161575.00± 10637.00 AUC _{(0- ∞)} hr*ng/mL 150081.26± 39104.00 88432.00± 24213.00 322800.00± 44567.00 154121.00± 28747.00 163878.00± 12089.00 MRT _(0-t) hr 99.62±6.26 74.20±13.83 96.50±15.90 76.40±12.70 97.50±9.67 MRT _{(0- ∞)} hr 104.89±7.58 77.40±13.46 101.00±19.2 77.8012.20 103.00±13.10 Table 8 Average pharmacokinetic parameters after single intravenous administration in cynomolgus monkeys PK parameters Unit Tirzepatide P016 C0 ng/mL 1167.70±159.90 1699.50±334.94 T1/2 hr 59.53±5.22 59.81±8.95 AUC _(0-t) hr*ng/mL 57554.64±11825..48 78169.25±20321.82 AUC _{(0- ∞)} hr*ng/mL 58529.59±11927.99 80032.34±21063.24 MRT _(0-t) hr 68.34±8.61 65.29±8.85 MRT _{(0- ∞)} hr 74.27±8.76 72.94±10.91 The PK test of the compound of the present invention injected subcutaneously in cynomolgus monkeys showed that the half-life of P014, P016 and P020 was equivalent to that of Tirzepatide, and the Tmax of P016 and P017 was delayed compared with Tirzepatide. The compound of the present invention modified with modified chains can achieve a plasma half-life equivalent to that of Tirzepatide, a dual-target product on the market, and the Tmax of P016 and P017 is about 2 times that of Tirzepatide, and the peak time of drug concentration is significantly delayed compared with Tirzepatide, which may have milder/less gastrointestinal reactions or a shorter titration time.

3. Pharmacodynamics

(I) Pharmacodynamics in db/db mice: To study the effect of the compound of the present invention on blood glucose in diabetic model mice (db/db mice). This study used a single subcutaneous administration of the compound to db/db mice to detect changes in blood glucose, food intake, and body weight to demonstrate the hypoglycemic effect and duration of the compound of the present invention, and to compare it with positive controls Tirzepatide and P001. In this study, male db/db mice aged 8-9 weeks were used. The db/db mice were housed in an independent ventilated cage IVC facility with controlled temperature (20-26°C) and humidity (40-70%), with a 12 h:12 h light/dark cycle, and free access to food and water. Blood was collected from the tip of the tail, and basal blood glucose was tested with a Roche blood glucose meter. The subjects were randomly divided into groups (n=6/group) according to the initial blood glucose and initial body weight, and each group had similar body weight and blood glucose. The compound of the present invention (10 nmol/kg) or the positive control Tirzepatide (10 nmol/kg) and P001 (10 nmol/kg) were dissolved in a solvent (PBS containing 0.1% Tween20, pH 7.2-7.4). After a single subcutaneous administration, the random blood glucose at the set time point (0-120 h, when the random blood glucose of the compound was no different from that of the Vehicle group, the random blood glucose of the compound was stopped), as well as the daily body weight and food intake were recorded. The data were analyzed using GraphPad Prism8. T-TEST was used to analyze the statistical differences between the groups. P < 0.05 was considered significant. Duration of hypoglycemic effect: The longest time when the random blood glucose of the compound was significantly different from that of the Vehicle group (P < 0.05). Table 9. Duration of hypoglycemic effect of the compound Group Compound Duration of hypoglycemic effect (h) Group 1 P001 32 Tirzepatide 56 P007 56 P008 72 P014 104 P019 56 Group 2 Tirzepatide 48 P013 72 P015 32 P016 104 P017 72 P018 48 P020 104 Compared with P001, the hypoglycemic effect of the compounds of the present invention lasts 24 hours or longer (P007, P008, P014, P019), even longer than 40 hours (P008, P014), and longer than 72 hours (P014) (see Table 9, Figure 1A, Figure 1B). Compared with Tirzepatide, the hypoglycemic effect of the compounds of the present invention lasts for a comparable or longer period of time (P007, P008, P013, P014, P016, P017, P018, P019, P020), and may even last for 16 hours or more (P008, P013, P014, P016, P017, P020), 24 hours or more (P013, P014, P016, P017, P020), 48 hours or more (P014, P016, P020), and 56 hours or more (P016, P020) (see Table 9). Compared with P001: Based on the test described in the above method, during the test period, in terms of hypoglycemic efficacy, the hypoglycemic effects of P008 (p < 0.01), P014 (p < 0.001) and P019 (p < 0.05) were significantly better than those of P001 (blood glucose AUC, P < 0.05, as shown in Figure 2A), and the blood glucose AUC inhibition rates were 2.3, 2.6 and 2.2 times that of P001, respectively; the hypoglycemic duration was 40 h, 72 h and 24 h longer than that of P001, respectively (Table 9). The hypoglycemic effect of P007 was equivalent to that of P001, but the hypoglycemic duration was 24 h longer than that of P001 (as shown in Figure 2A and Table 9). Compared with Tirzepatide: Compared with Tirzepatide, P014 (p < 0.05, Figure 2B), P016 (p < 0.05, Figure 2C) and P020 (p < 0.05, Figure 2C) had stronger hypoglycemic effects (blood glucose AUC, p < 0.05), and the blood glucose AUC inhibition rates were 1.7, 2.2 and 2.0 times that of Tirzepatide, respectively; moreover, they also showed significantly longer duration of drug effect (48 h, 56 h and 56 h longer than Tirzepatide, respectively, Table 9). The hypoglycemic effects of P013 and P017 were comparable to those of Tirzepatide (Figure 2C), but the duration of drug effect was 24 h longer than that of Tirzepatide (Table 9). Unexpectedly, compared with P001, the acylated GLP-1/GIP agonist of the present invention significantly improves the hypoglycemic effect while achieving a long-lasting effect. The compounds of the present invention with different forms of modified chains can achieve a stronger and more lasting hypoglycemic effect than P001, and some of the compounds of the present invention show a stronger and more lasting hypoglycemic effect than Tirzepatide. And in combination with the target binding and agonistic activity of the compounds of the present invention on human GLP-1 and GIP receptors, it was unexpectedly found that the target binding and agonistic activity of the compounds of the present invention on the human GLP-1 receptor was significantly reduced compared with P001, and the target binding and agonistic activity of the human GIP receptor was also significantly reduced, but the hypoglycemic effect of the compounds of the present invention was significantly better than that of P001.

(II) Multiple dosing experiment in db/db mice In this study, the compounds of the present invention were administered subcutaneously to db/db mice multiple times, and the changes in blood glucose in mice were detected to further illustrate the hypoglycemic effect of the compounds of the present invention, and compared with the positive control Tirzepatide. In this study, male db/db mice aged 7-8 weeks were used. The db/db mice were placed in an IVC facility with controlled temperature (20-26°C) and humidity (40-70%), controlled by a 12 h:12 h light/dark cycle, and free access to food and water. Blood was collected from the tip of the tail, and basal blood glucose was detected with a Roche blood glucose meter. The mice were randomly divided into groups (n=6/group) according to the initial blood glucose and initial body weight, and each group had similar body weight and blood glucose. The compound of the present invention (3, 10, 30 nmol/kg) or the positive control Tirzepatide (3, 10, 30 nmol/kg) was dissolved in a solvent (PBS containing 0.1% Tween20, pH 7.2-7.4). The drug was administered subcutaneously once every 3 days for four weeks. The drug was administered subcutaneously on days 1, 4, 7, 10, 13, 16, 19, 22 and 25. Random blood glucose was recorded every 3 days throughout the study. Fasting blood glucose was tested on day 28, and then blood was collected by orbital bleeding. After orbital bleeding, the animals were killed, and the liver was removed and weighed. After 1 to 2 hours of the collected blood, the serum was separated by centrifugation at 3000 rpm for 10 minutes. Biochemical indicators such as triglyceride (TG), alanine aminotransferase (ALT), and total bilirubin (TBIL) were detected using a biochemical instrument. The data were statistically analyzed using GraphPad Prism8, and the statistical differences between the groups were analyzed using T-TEST. P < 0.05 was considered to be significant. In the experiments conducted as described above, the compounds of the present invention, such as P014, P016, P017 and P020, showed better blood glucose inhibition than Tirzepatide at a dose of 3 nmol/kg; at a dose of 30 nmol/kg, the efficacy of P016 was superior to that of Tirzepatide (see Figures 3A-3D, Table 10). P016, P017 and P020 could significantly reduce the liver weight of db/db mice at multiple doses, while Tirzepatide did not reduce the liver weight at all three experimental doses. The compounds of the present invention, such as P014, P016, P017, P020 and Tirzepatide, can effectively reduce serum TG in db/db mice (except for the P017 and Tirzepatide 3 nmol/kg groups), and the efficacy of some dose groups of P016 and P020 is superior to that of Tirzepatide (see Table 11). In addition, P014 and P016 can also reduce serum TBIL and/or ALT. Table 10. Fasting blood glucose and blood glucose AUC of db/db mice Group D28 fasting blood glucose (mmol/L) AUC _{4-25 day} Glu (mmol/L·day) Vehicle 18.34±3.45 493.71±40.3 P014 3 nmol/kg 10.22±1.58* ^## 294±40.43** ^# P014 10 nmol/kg 10.72±1.75 214.4±33.73***# P014 30 nmol/kg 13.38±3.38 190.62±26.47*** P016 3 nmol/kg 10.42±1.46* ^## 256.38±22*** ^### P016 10 nmol/kg 12.5±2.19 249.25±41.42** P016 30 nmol/kg 6.05±0.41** ^## 163.98±11.57**** ^# P017 3 nmol/kg 11.68±1.7 ^# 303.78±33.24** ^# P017 10 nmol/kg 11.95±2.51 249.65±47.65** P017 30 nmol/kg 10.08±0.79* 294.96±11.96** P020 3 nmol/kg 9.6±1.32* ^## 268.28±24.64*** ^# P020 10 nmol/kg 10.7±1.62 290.78±27.25** P020 30 nmol/kg 8.22±1.07* 184.18±22.44**** TZP 3 nmol/kg 17.35±1.21 419.85±26.42 TZP 10 nmol/kg 13.82±1.99 355.68±30.8* TZP 30 nmol/kg 10.88±1.2 252.7±32.81** T-Test, *p＜0.05, **p＜0.01, ***p＜0.001, ****p＜0.0001 compared with vehicle group; #p＜0.05, ##p＜0.01, ###p＜0.001 compared with Tirzepatide (TZP) same dose group. The results are expressed as Mean±SEM of 6 mice. Table 11. Liver weight and serum TG of db/db mice Group Liver weight (g) Serum TG (mmol/L) Vehicle 2.61±0.19 1.87±0.13 P014 3 nmol/kg 2.26±0.18 1.39±0.07* P014 10 nmol/kg 2.1±0.14 1.29±0.03** P014 30 nmol/kg 1.89±0.31 1.14±0.18* P016 3 nmol/kg 2.24±0.1 ^# 1.21±0.04*** ^# P016 10 nmol/kg 1.98±0.12* 1.22±0.24* P016 30 nmol/kg 1.78±0.08** 1.2±0.05*** P017 3 nmol/kg 2.14±0.08* ^## 1.58±0.17 P017 10 nmol/kg 2.11±0.09* 1.19±0.05*** P017 30 nmol/kg 2.35±0.09 1.09±0.06*** P020 3 nmol/kg 2.03±0.09* ^## 1.37±0.12* P020 10 nmol/kg 2.09±0.1* 1.07±0.03*** ^# P020 30 nmol/kg 1.83±0.16* 1.2±0.07** TZP 3 nmol/kg 2.49±0.05 1.52±0.11 TZP 10 nmol/kg 2.31±0.17 1.21±0.04** TZP 30 nmol/kg 2.07±0.15 1.18±0.12** T-Test, *p＜0.05, **p＜0.01, ***p＜0.001 compared with vehicle group; #p＜0.05, ##p＜0.01 compared with Tirzepatide (TPZ) same dose group. The results are expressed as Mean±SEM of 6 mice. Table 12. Serum ALT and TBIL of db/db mice Group Serum ALT (U/L) Serum TBIL (μmol/L) Vehicle 189.55±40.92 1.43±0.3 P014 3 nmol/kg 111.24±13.23 0.58±0.11* ^## P014 10 nmol/kg 89.9±8.66* 0.58±0.15* P014 30 nmol/kg 106.47±14.3 0.36±0.15* ^## P016 3 nmol/kg 137.93±34.59 0.82±0.13 P016 10 nmol/kg 128.94±16.93 1.22±0.28 P016 30 nmol/kg 97.7±12.36* 0.73±0.16 P017 3 nmol/kg 105.16±7.1 0.88±0.26 P017 10 nmol/kg 141.65±12.72 1.59±0.07 P017 30 nmol/kg 176.48±51.6 1.11±0.1 P020 3 nmol/kg 214.56±43.83 1.54±0.25 P020 10 nmol/kg 113.48±24.18 0.91±0.05 P020 30 nmol/kg 112.47±16.69 0.95±0.28 TZP 3 nmol/kg 146.03±19.67 1.54±0.17 TZP 10 nmol/kg 138.66±31.26 1.2±0.3 TZP 30 nmol/kg 112.48±14.55 0.91±0.04 T-Test, *p＜0.05 compared with vehicle group; #p＜0.05, ##p＜0.01 compared with Tirzepatide (TPZ) group with the same dose. The results are expressed as Mean±SEM of 6 mice. In summary, under this experimental system, the compounds of the present invention can effectively reduce the blood glucose of db/db mice by subcutaneous injection for 4 weeks, and the effective dose (≤3 nmol/kg) is 1/3 of Tirzepatide (10 nmol/kg), and the maximum efficacy (30 nmol/kg) of some compounds of the present invention, such as P016, is stronger than Tirzepatide. In addition, some compounds of the present invention show a reduction in liver weight and a protective effect on the liver in db/db mice, while Tirzepatide has no similar effect.

(III) Repeated drug administration for 2 weeks in male rats 1. Experimental design 77 SD male rats (SPF grade) were selected and randomly divided into 10 groups according to body weight. Group 1 was the solvent control group, with 5 animals; Groups 2 to 4 were the commercial drug control groups (Tirzepatide), with 8 animals in each group, including 5 animals in the main test and 3 animals in the toxicity test, and the drug dosages were 1, 3, and 10 mg/kg respectively; Groups 5 to 7 were the test substance group 1, and groups 8 to 10 were the test substance group 2, with 8 animals in each group, including 5 animals in the main test and 3 animals in the toxicity test, and the drug dosages were 1, 3, and 10 mg/kg respectively. All the above groups were administrated twice a week for 2 consecutive weeks (administration days were: D1, 5, 8, 12, and 15). During the trial, all groups of animals were subjected to cage observation, detailed clinical observation, food consumption measurement, body weight measurement, blood biochemistry, hematology index measurement, gross anatomical observation, organ weighing, and toxicokinetics study. 2. Data Statistics The following table details the combination used for statistical comparison. See the table below: Statistical Method Table Control group Comparison Group 1 2, 3, 4, 5, 6, 7, 8, 9, 10 Raw data for each time period will be tabulated, and means and standard deviations and/or category changes will be calculated by group and sex for each endpoint. For each endpoint, the treated group will be compared to the control group as described below. Endpoint data will be log-transformed before specific analyses are performed as required. Statistical Analysis Table Indicators Analytical method Food consumption Body weight Hematology (excluding leukocyte count) Coagulation Serum biochemistry Organ weight Absolute weight relative to body weight and brain organ coefficients Comparison between groups White blood cell count White blood cell count White blood cell count Logarithmic transformation/comparison between groups 3. Experimental results Body weight/weight gain: After the first administration, weight loss was observed in the following dosage groups in each dosage group: Tirzepatide ≥ 3 mg/kg; P014 and P016 ≥ 1 mg/kg. As the administration continued, weight recovered to some extent. At the end of the administration, weight of all groups was increasing, but weight gain was less than that of the control group, showing a significant dose-related relationship. Food intake: Throughout the entire trial, a dose-related decrease in food intake was observed in each dosage group (Tirzepatide, P014 and P016). Blood biochemistry: Amylase and triglyceride levels decreased. Hematology: Reticulocyte levels were slightly decreased. Gross autopsy: No abnormalities were found in any dosage group (Tirzepatide, P014 and P016). Organ weight: The weight of heart, liver, spleen, etc. in each dosing group (Tirzepatide, P014 and P016) decreased, which was not much different from the vehicle control group, which may be related to weight loss and is considered to be related to pharmacological effects. Toxicokinetics: _Cmax and AUC of Tirzepatide, P014 and P016 groups increased with dose in a form close to dose proportion; the drug was given twice a week for 2 consecutive weeks, and no obvious accumulation was observed in each dose group of TZP, P014 and P016. Each dosing group (Tirzepatide, P014 and P016) was administered twice a week, and the MTD for 2 consecutive weeks was ≥10 mg/kg. The C _max and AUC _last after the last dose and the safety window of the trial were as follows: Table 13. Safety window of each dosing group Experimental Project Tirzepatide P014 P016 MTD 2077 nmol/kg (2-week trial MTD) 1980 nmol/kg (2-week trial MTD) 1924 nmol/kg (2-week trial MTD) Efficacy Dose 10 nmol/kg 3 nmol/kg 3 nmol/kg Safety window (depending on dosage) 416 1319 1282 Safety window (based on exposure AUC _last ) 130 596 920

In summary, the toxicity properties of P016 and P014 are similar to those of Tirzepatide, but they have a higher safety window (based on exposure, the safety window of P014 is more than 4 times that of Tirzepatide, while the safety window of P016 is more than 7 times that of Tirzepatide).

In addition, relative to the safety window of P001, the safety window of the compounds of the present invention is 10 times, or 20 times, or 30 times, or 40 times, or 50 times, or 60 times, or 70 times, or 80 times, or 90 times larger than that of P001, especially P014 and P016.

At the same time, at the same exposure, the effect of the compound of the present invention on HR is lower than that of Tirepatide, the heart rate increase of P016 is lower than that of Tirepatide, and there is no sustained heart rate increase of more than 30%. At the same exposure, the heart rate recovery of P016 is faster than that of TZP (the recovery time of P016 is between 72 and 120 hours, and the recovery time of TZP is greater than 120 hours).

The above is a detailed introduction to the peptide compounds and applications provided by the present invention.

The principles and implementation methods of the present invention are described in detail using specific examples. The above examples are only used to help understand the method and its central idea of the present invention. It should be pointed out that ordinary technicians in this field can make several improvements and modifications to the present invention without departing from the principles of the present invention. These improvements and modifications also fall within the scope of protection of the patent application of the present invention. Amino acid sequence: Name Serial number sequence P001 SEQ ID NO:1 YX ₁ -EGTX ₂ -TSDYX ₃ -IX ₄ -LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein X ₁ is Aib; X ₂ is αMePhe; X ₃ is Aib; X ₄ is Aib; and the C-terminal amino acid is amidated to form a C-terminal primary amide (SEQ ID NO: 1) P007 SEQ ID NO:7 _YX1 - _EGTX2 -TSDYX3- _{IX4 -} LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at position 24 is chemically modified by binding ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _2- (γ-Glu) ₁ -CO-( _CH2 ) ₁₆ - _CO2H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO:7). P008 SEQ ID NO:8 _YX1 - _EGTX2 -TSDYX3- _{IX4 -} LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at position 28 is chemically modified by binding ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _2- (γ-Glu) ₁ -CO-( _CH2 ) ₁₆ - _CO2H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO:8). P013 SEQ ID NO:13 _YX1 - _EGTX2 -TSDYX3- _{IX4 -} LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at position 24 is chemically modified by binding ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _2- (γ-Glu) ₁ -CO-( _CH2 ) ₁₈ - _CO2H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 13). P014 SEQ ID NO:14 _YX1 - _EGTX2 -TSDYX3- _{IX4 -} LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at position 24 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _2- (γ-Glu) ₁ -CO-( _CH2 ) ₂₀ - _CO2H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 14). P015 SEQ ID NO:15 YX ₁ -EGTX ₂ -TSDYX ₃ -IX ₄ -LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein X ₁ is Aib; X ₂ is αMePhe; X ₃ is Aib; X ₄ is Aib; K at position 24 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) ₂ -(γ-Glu) ₃ -CO-(CH ₂ ) ₁₆ -CO ₂ H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 15) P016 SEQ ID NO:16 YX ₁ -EGTX ₂ -TSDYX ₃ -IX ₄ -LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein X ₁ is Aib; X ₂ is αMePhe; X ₃ is Aib; X ₄ is Aib; K at position 24 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) ₃ -(γ-Glu) ₁ -CO-(CH ₂ ) ₂₀ -CO ₂ H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 16) P017 SEQ ID NO:17 YX ₁ -EGTX ₂ -TSDYX ₃ -IX ₄ -LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein X ₁ is Aib; X ₂ is αMePhe; X ₃ is Aib; X ₄ is Aib; K at position 28 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) ₂ -(γ-Glu) ₁ -CO-(CH ₂ ) ₂₀ -CO ₂ H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 17) P018 SEQ ID NO:18 YX ₁ -EGTX ₂ -TSDYX ₃ -IX ₄ -LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein X ₁ is Aib; X ₂ is αMePhe; X ₃ is Aib; X ₄ is Aib; K at position 28 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) ₂ -(γ-Glu) ₃ -CO-(CH ₂ ) ₁₆ -CO ₂ H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 18) P019 SEQ ID NO:19 YX ₁ -EGTX ₂ -TSDYX ₃ -IX ₄ -LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein X ₁ is Aib; X ₂ is αMePhe; X ₃ is Aib; X ₄ is Aib; K at position 28 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) ₃ -(γ-Glu) ₁ -CO-(CH ₂ ) ₁₆ -CO ₂ H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 19) P020 SEQ ID NO:20 YX ₁ -EGTX ₂ -TSDYX ₃ -IX ₄ -LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein X ₁ is Aib; X ₂ is αMePhe; X ₃ is Aib; X ₄ is Aib; K at position 28 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) ₃ -(γ-Glu) ₁ -CO-(CH ₂ ) ₂₀ -CO ₂ H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 20)

Figures 1A-1B show the changes in blood glucose levels in db/db mice after drug administration. Figure 1A shows the changes in blood glucose levels in db/db mice after a single subcutaneous administration of P007, P008, P014, P019, Tirzepatide, and P001 at 10 nmol/kg; Figure 1B shows the changes in blood glucose levels in db/db mice after a single subcutaneous administration of P013, P015, P016, P017, P018, P020, and Tirzepatide at 10 nmol/kg.

Figures 2A-2C show the blood glucose AUC of db/db mice after administration. Figure 2A shows the AUC of P007, P008, P014, P019, Tirzepatide and P001 at 0-48 h after a single subcutaneous administration of 10 nmol/kg; Figure 2B shows the AUC of P007, P008, P014, P019 and Tirzepatide at 0-72 h after a single subcutaneous administration of 10 nmol/kg; Figure 2C shows the AUC of P013, P015, P016, P017, P018, P020 and Tirzepatide at 0-56 h after a single subcutaneous administration of 10 nmol/kg. In Figure 2A, T-Test, *p＜0.05, **p＜0.01, ***p＜0.001 compared with Vehicle; §p＜0.05, §§p＜0.01, §§§p＜0.001 compared with P001. In Figure 2B, T-Test, *p＜0.05, **p＜0.01, ***p＜0.001 compared with Vehicle; #p＜0.05, ##p＜0.01 compared with Tirzepatide. In Figure 2C, T-Test, *p＜0.05, **p＜0.01, ***p＜0.001 compared with Vehicle; #p＜0.05 compared with Tirzepatide.

Figures 3A-3D show the changes in blood glucose levels of db/db mice after multiple dosing. Figure 3A compares the changes in blood glucose levels of db/db mice after multiple subcutaneous dosing with P014 and Tirzepatide; Figure 3B compares the changes in blood glucose levels of db/db mice after multiple subcutaneous dosing with P016 and Tirzepatide; Figure 3C compares the changes in blood glucose levels of db/db mice after multiple subcutaneous dosing with P017 and Tirzepatide; Figure 3D compares the changes in blood glucose levels of db/db mice after multiple subcutaneous dosing with P020 and Tirzepatide.

The present invention also includes new intermediates and methods that can be used to synthesize the compounds of the present invention or their pharmaceutically acceptable salts. The intermediates and compounds of the present invention can be prepared by a variety of methods known in the art. In particular, the methods using chemical synthesis are illustrated in the following examples. The specific synthetic steps of each described approach can be combined in different ways to prepare the compounds of the present invention or their salts. Reagents and raw materials are easily available to ordinary technicians in this field. It should be understood that these examples are not intended to limit the scope of the present invention in any way. The mass spectrometer uses an Agilent 1260/6110 liquid chromatography mass spectrometer with a scanning range of 100-1500.

TW202411244A_112126024_SEQL.xml

Claims (17)

A compound of the following formula (AI) or a pharmaceutically acceptable salt thereof, YX ₁ -EGTX ₂ -TSDY-A11-A12-A13-LDK-A17-AQ-A20-EFVKWLLK-A29-GPSSGAPPPSK Formula (AI); wherein X ₁ is Aib; X ₂ is αMePhe; A11 is Aib or Ala; A12 is Ala, Ile, Lys, Phe or Pya(4); A13 is Aib, Cha, Leu, αMePhe or αMeTyr; A17 is Gln or Ile; A20 is Ala or Ser; A29 is Gln or Gly, 1 K, 2 K, 3 K or 4 K positions are selected from among positions 16, 24, 28 and 40, by using ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -Z is chemically modified by binding to the ε-amine group of the K side chain, wherein each a is independently an integer from 0 to 5, each b is independently an integer from 0 to 5, each c is independently an integer from 10 to 24, and Z is independently selected from -CH ₃ , carboxylic acid or carboxylic acid bioisostere, phosphate/ester or sulfonate/ester. A compound or a pharmaceutically acceptable salt thereof as claimed in claim 1, wherein said A11 is Aib; or/and A12 is Ile; or/and A13 is Aib; or/and A17 is Gln; or/and A20 is Ala; or/and A29 is Gly; or/and a is independently selected from an integer of 1, 2, 3, 4 or 5, b is independently selected from an integer of 1, 2, 3, 4 or 5, c is independently selected from an integer of 12 to 22; further c is independently selected from 14, 16, 18 or 20. A compound of the following formula (I) or a pharmaceutically acceptable salt thereof, _YX1 - _EGTX2 - _TSDYX3 - _IX4- LDKQAQAEFVKWLLKGGPSSG-APPPSK Formula (I); wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; 1, 2, 3 or 4 K positions are selected from the 16th, 24th, 28th and 40th positions, by using ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a- (γ-Glu) _b -CO-( _CH2 ) _c -Z is chemically modified by binding to the ε-amine group of the K side chain, wherein each a is independently an integer from 0 to 5, each b is independently an integer from 0 to 5, each c is independently an integer from 10 to 24, wherein Z is independently selected from -CH ₃ , carboxylic acid or carboxylic acid bioisostere, phosphate/ester or sulfonate/ester; and the C-terminal amino acid is amidated to form a C-terminal primary amide. The compound of claim 3 or a pharmaceutically acceptable salt thereof, wherein the compound is: YX ₁ -EGTX ₂ -TSDYX ₃ -IX ₄ -LDKQAQAEFVKWLLKGGPSSG-APPPSK Formula (I), or a pharmaceutically acceptable salt thereof, wherein X ₁ is Aib; X ₂ is αMePhe; X ₃ is Aib; X ₄ is Aib; one or two K positions selected from positions 16, 24, 28, and 40 are chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -Z to the ε-amine group of the K side chain, wherein Z is independently selected from -CH ₃ , carboxylic acid or carboxylic acid bioisostere, phosphate/ester or sulfonate/ester, preferably -CO ₂ H, each a is independently an integer from 0 to 5, each b is independently an integer from 0 to 5, and each c is independently an integer from 10 to 22; and the C-terminal amino acid is amidated to form a C-terminal primary amide. The compound or a pharmaceutically acceptable salt thereof of claim 4, wherein the 16-position K is chemically modified by binding to the ε-amine group of the K side chain with ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -CO ₂ H; or the 24-position K is chemically modified by binding to the ε-amine group of the K side chain with ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -CO ₂ H; or the 28-position K is chemically modified by binding to the ε-amine group of the K side chain with ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -CO ₂ H is chemically modified by binding to the ε-amine group of the K side chain; or at the 40-position of K, it is chemically modified by binding to the ε-amine group of the K side chain with ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -CO ₂ H. The compound or pharmaceutically acceptable salt thereof of claim 4, wherein the K at positions 24 and 28 is chemically modified by binding to the ε-amine group of the K side chain with ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -CO ₂ H to perform chemical modification of two modification chains; or the K at positions 16 and 24 is chemically modified by binding to the ε-amine group of the K side chain with ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -CO ₂ H to perform chemical modification of two modification chains; or the K at positions 16 and 28 is chemically modified by binding to the ε-amine group of the K side chain with ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -CO ₂ H is bound to the ε-amine group of the K side chain to chemically modify the two modification chains; or K at positions 16 and 40 is chemically modified by binding to the ε-amine group of the K side chain with ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -CO ₂ H; or K at positions 24 and 40 is chemically modified by binding to the ε-amine group of the K side chain with ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -CO ₂ H is bound to the ε-amine group of the K side chain to perform chemical modification of two modification chains; or K at positions 28 and 40 is chemically modified by binding ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a -(γ-Glu) _b -CO-(CH ₂ ) _c -CO ₂ H to the ε-amine group of the K side chain. The compound or pharmaceutically acceptable salt thereof of any one of claims 3 to 6, wherein a is independently selected from 1, 2, 3, 4 or 5, b is independently selected from 1, 2, 3, 4 or 5, c is independently selected from 12 to 22; further c is independently selected from 14, 16, 18 or 20; and/or Z is -CO ₂ H. The compound of claim 4 or a pharmaceutically acceptable salt thereof, wherein the compound is _YX1 - _EGTX2 -TSDYX3- _{IX4 -} LDKQAQAEFVKWLLKGGPSSG-APPPSK of formula (I); wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at position 24 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a- (γ-Glu) _b -CO-( _CH2 ) _c - _CO2H to the ε-amine group of the K side chain, wherein a is independently selected from an integer from 0 to 5, b is independently selected from an integer from 0 to 5, and c is independently selected from an integer from 10 to 24; and the C-terminal amino acid is amidated to form a C-terminal primary amide. The compound of claim 4 or a pharmaceutically acceptable salt thereof, wherein the compound is _YX1 - _EGTX2 -TSDYX3- _{IX4 -} LDKQAQAEFVKWLLKGGPSSG-APPPSK of formula (I); wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at position 28 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _a- (γ-Glu) _b -CO-( _CH2 ) _c - _CO2H to the ε-amine group of the K side chain, wherein a is independently selected from an integer from 0 to 5, b is independently selected from an integer from 0 to 5, and c is independently selected from an integer from 10 to 24; and the C-terminal amino acid is amidated to form a C-terminal primary amide. A compound or a pharmaceutically acceptable salt thereof as claimed in any one of claims 8 to 9, wherein said a is independently selected from an integer of 1, 2, 3, 4 or 5, b is independently selected from an integer of 1, 2, 3, 4 or 5, and c is independently selected from an integer of 12 to 22; further, c is independently selected from 14, 16, 18 or 20. The compound of claim 4 or a pharmaceutically acceptable salt thereof, wherein the compound is YX ₁ -EGTX ₂ -TSDYX ₃ -IX ₄ -LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein X ₁ is Aib; X ₂ is αMePhe; X ₃ is Aib; X ₄ is Aib; K at position 24 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) ₂ -(γ-Glu) ₁ -CO-(CH ₂ ) ₁₆ -CO ₂ H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to form a C-terminal primary amide (SEQ ID NO: 7); or YX ₁ -EGTX ₂ -TSDYX ₃ -IX ₄ -LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein X _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at position 24 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _2- (γ-Glu) ₁ -CO-( _CH2 ) ₁₈ -CO2H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO:13); or _YX1 - _EGTX2-TSDYX3 - _{IX4 -} LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at position 24 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl)2-(γ-Glu)1-CO-(CH2) ₁₈ -CO2H to the ε-amine group of the K side chain; and the _C -terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO:13). -(γ-Glu) ₁ -CO-(CH ₂ ) ₂₀ -CO ₂ H is chemically modified to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 14); or YX ₁ -EGTX ₂ -TSDYX ₃ -IX ₄ -LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein X ₁ is Aib; X ₂ is αMePhe; X ₃ is Aib; X ₄ is Aib; and the K at position 24 is modified by using ([2-(2-amino-ethoxy)-ethoxy]-acetyl) ₂ -(γ-Glu) ₃ -CO-(CH ₂ ) ₁₆ -CO ₂ H is chemically modified by binding to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 15); or YX ₁ -EGTX ₂ -TSDYX ₃ -IX ₄ -LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein X ₁ is Aib; X ₂ is αMePhe; X ₃ is Aib; X ₄ is Aib; K at position 24 is chemically modified by binding to the ε-amine group of the K side chain with ([2-(2-amino-ethoxy)-ethoxy]-acetyl) ₃ -(γ-Glu) ₁ -CO-(CH ₂ ) ₂₀ -CO ₂ H; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 16); or YX 1 -EGTX 2 -TSDYX 3 -IX 4 -LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein X 1 is Aib; X 2 is αMePhe; X 3 is Aib; X 4 is Aib; K at position 24 is chemically modified by binding to the ε-amine group of the K side chain with ([2-(2-amino-ethoxy)-ethoxy]-acetyl) 3 -(γ-Glu) ₁ -CO-(CH 2 ) 20 -CO ₂ -TSDYX ₃ -IX ₄ -LDKQAQAEFVKWLLKGGPSSG-APPPSK ； wherein X ₁ is Aib; X ₂ is αMePhe; X ₃ is Aib; X ₄ is Aib; K at position 28 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) ₂ -(γ-Glu) ₁ -CO-(CH ₂ ) ₁₆ -CO ₂ H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 8); or YX ₁ -EGTX ₂ -TSDYX ₃ -IX ₄ -LDKQAQAEFVKWLLKGGPSSG-APPPSK ； wherein X ₁ is Aib; X ₂ is αMePhe; X ₃ is Aib; X 4 is Aib; wherein X1 is Aib; X2 is αMePhe; X3 is Aib; _X4 is Aib; K at position 28 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _2- (γ-Glu) ₁ -CO-( _CH2 ) ₂₀ - _CO2H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO:17); or _YX1 - _EGTX2 - _TSDYX3 - _IX4 -LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein _X1 is Aib; _X2 is αMePhe; _X3 is Aib; _X4 is Aib; K at position 28 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) _2- (γ-Glu) ₃ -CO-( _CH2 ) ₁₆ -CO ₂ H is chemically modified by conjugating to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 18); or YX ₁ -EGTX ₂ -TSDYX ₃ -IX ₄ -LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein X ₁ is Aib; X ₂ is αMePhe; X ₃ is Aib; X ₄ is Aib; K at position 28 is chemically modified by conjugating to the ε-amine group of the K side chain with ([2-(2-amino-ethoxy)-ethoxy]-acetyl) ₃ -(γ-Glu) ₁ -CO-(CH 2 ) 16 -CO ₂ H; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 19); or YX 1 -EGTX 2 -TSDYX 3 -IX 4 -LDKQAQAEFVKWLLKGGPSSG-APPPSK; wherein X 1 is Aib; X 2 is αMePhe; X 3 is Aib; X ₄ is Aib; K at position 28 is chemically modified by conjugating to the ε-amine group of the K side chain with ([2-(2-amino-ethoxy)-ethoxy]-acetyl) 3 -(γ-Glu) 1 -CO-(CH ₂ ) ₁₆ -CO 2 H; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 19); ₂ -TSDYX ₃ -IX ₄ -LDKQAQAEFVKWLLKGGPSSG-APPPSK ; wherein X ₁ is Aib; X ₂ is αMePhe; X ₃ is Aib; X ₄ is Aib; K at position 28 is chemically modified by conjugating ([2-(2-amino-ethoxy)-ethoxy]-acetyl) ₃ -(γ-Glu) ₁ -CO-(CH ₂ ) ₂₀ -CO ₂ H to the ε-amine group of the K side chain; and the C-terminal amino acid is amidated to a C-terminal primary amide (SEQ ID NO: 20). The compound of claim 4 or a pharmaceutically acceptable salt thereof, wherein the compound is , , , , , , , , , , or a pharmaceutically acceptable salt thereof. A pharmaceutical composition comprising a compound according to any one of claims 1 to 12 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier, diluent or excipient. A use of a compound according to any one of claims 1 to 12 or a pharmaceutically acceptable salt thereof, or a composition according to claim 13, for preparing a medicament for treating or preventing the following diseases: hyperglycemia, impaired glucose tolerance, type I diabetes, type II diabetes, obesity, hypertension, syndrome X, dyslipidemia, cognitive impairment, atherosclerosis, myocardial infarction, coronary heart disease and other cardiovascular diseases, stroke, inflammatory bowel syndrome, dyspepsia and gastric ulcer. A use of the compound of any one of claims 1 to 12 or a pharmaceutically acceptable salt thereof, or the composition of claim 13, for preparing a medicament for delaying or preventing the progression of type II diabetes. A use of a compound of any one of claims 1 to 12 or a pharmaceutically acceptable salt thereof, or a composition of claim 13, for preparing a medicament for reducing food intake, reducing β-cell apoptosis, increasing β-cell function and β-cell mass and/or restoring β-cell glucose sensitivity. A use of a compound according to any one of claims 1 to 12 or a pharmaceutically acceptable salt thereof, or a composition according to claim 13, for preparing a medicament for treating type II diabetes.